Ras inhibitors

ABSTRACT

The disclosure features macrocyclic compounds, and pharmaceutical compositions and protein complexes thereof, capable of inhibiting Ras proteins, and their uses in the treatment of cancers.

CROSS-REFERENCE TO RELATED APPLICATIONS

The present application claims the benefit of priority to U.S.Application No. 63/078,802, filed on Sep. 15, 2020; U.S. Application No.63/129,231, filed on Dec. 22, 2020; U.S. Application No. 63/184,412,filed on May 5, 2021; and U.S. Application No. 63/192,775, filed on May25, 2021, all of which are hereby incorporated by reference in theirentirety.

SEQUENCE LISTING

The instant application contains a Sequence Listing which has beensubmitted electronically in ASCII format and is hereby incorporated byreference in its entirety. Said ASCII copy, created on Sep. 14, 2021, isnamed 51432-009005_Sequence_Listing_9_14_21_ST25 and is 1,189 bytes insize.

BACKGROUND

The vast majority of small molecule drugs act by binding a functionallyimportant pocket on a target protein, thereby modulating the activity ofthat protein. For example, cholesterol-lowering drugs known as statinsbind the enzyme active site of HMG-CoA reductase, thus preventing theenzyme from engaging with its substrates. The fact that many suchdrug/target interacting pairs are known may have misled some intobelieving that a small molecule modulator could be discovered for most,if not all, proteins provided a reasonable amount of time, effort, andresources. This is far from the case. Current estimates are that onlyabout 10% of all human proteins are targetable by small molecules.Bojadzic and Buchwald, Curr Top Med Chem 18: 674-699 (2019). The other90% are currently considered refractory or intractable towardabove-mentioned small molecule drug discovery. Such targets are commonlyreferred to as “undruggable.” These undruggable targets include a vastand largely untapped reservoir of medically important human proteins.Thus, there exists a great deal of interest in discovering new molecularmodalities capable of modulating the function of such undruggabletargets.

It has been well established in literature that Ras proteins (K-Ras,H-Ras and N-Ras) play an essential role in various human cancers and aretherefore appropriate targets for anticancer therapy. Indeed, mutationsin Ras proteins account for approximately 30% of ail human cancers inthe United States, many of which are fatal. Dysregulation of Rasproteins by activating mutations, overexpression or upstream activationis common in human tumors, and activating mutations in Ras arefrequently found in human cancer. For example, activating mutations atcodon 12 in Ras proteins function by inhibiting both GTPase-activatingprotein (GAP)-dependent and intrinsic hydrolysis rates of GTP,significantly skewing the population of Ras mutant proteins to the “on”(GTP-bound) state (Ras(ON)), leading to oncogenic MAPK signaling.Notably, Ras exhibits a picomolar affinity for GTP, enabling Ras to beactivated even in the presence of low concentrations of this nucleotide.Mutations at codons 13 (e.g., G13D) and 61 (e.g., Q61K) of Ras are alsoresponsible for oncogenic activity in some cancers.

Despite extensive drug discovery efforts against Ras during the lastseveral decades, a drug directly targeting the “on” form of Ras is stillnot approved. Additional efforts are needed to uncover additionalmedicines for cancers driven by the various Ras mutations.

SUMMARY

Provided herein are Ras inhibitors. These Ras inhibitors target, thatis, selectively bind to or inhibit, Ras(ON) (e.g., selective over theGDP-bound, inactive state of Ras). The approach described herein entailsformation of a high affinity three-component complex between a syntheticligand and two intracellular proteins which do not interact under normalphysiological conditions: the target protein of interest (e.g., Ras),and a widely expressed cytosolic chaperone (presenter protein) in thecell (e.g., cyclophilin A). More specifically, in some embodiments, theinhibitors of Ras described herein induce a new binding pocket in Ras bydriving formation of a high affinity tri-complex between the Ras proteinand the widely expressed cytosolic chaperone, cyclophilin A (CYPA).Without being bound by theory, the inventors believe that one way theinhibitory effect on Ras is effected by compounds of the invention andthe complexes they form is by steric occlusion of the interaction sitebetween Ras and downstream effector molecules, such as RAF and PI3K,which are required for propagating the oncogenic signal.

As such, in some embodiments, the disclosure features a compound, orpharmaceutically acceptable salt thereof, of structural Formula Ia:

wherein A is optionally substituted 3 to 6-membered cycloalkylene,optionally substituted 3 to 6-membered heterocycloalkylene, optionallysubstituted 6-membered arylene, optionally substituted 5 to 6-memberedheteroarylene, optionally substituted C₂-C₄ alkylene, or optionallysubstituted C₂-C₄ alkenylene;

Y is

W is hydrogen, C₁-C₄ alkyl, optionally substituted C₁-C₃ heteroalkyl,optionally substituted 3 to 10-membered heterocycloalkyl, optionallysubstituted 3 to 10-membered cycloalkyl, optionally substituted 6 to10-membered aryl, or optionally substituted 5 to 10-membered heteroaryl;

X¹ and X⁴ are each, independently, CH₂ or NH;

R¹ is optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 15-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 6-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl;

R² is hydrogen, optionally substituted C₁-C₆ alkyl, optionallysubstituted C₂-C₆ alkenyl, optionally substituted C₂-C₆ alkynyl,optionally substituted 3 to 6-membered cycloalkyl, optionallysubstituted 3 to 7-membered heterocycloalkyl, optionally substituted6-membered aryl, optionally substituted 5 or 6-membered heteroaryl; and

R¹⁰ is hydrogen, hydroxy, optionally substituted C₁-C₃ alkyl, oroptionally substituted C₁-C₆ heteroalkyl. In some embodiments, R¹⁰ ishydrogen.

Also provided are pharmaceutical compositions comprising a compound ofFormula Ia, or a pharmaceutically acceptable salt thereof, and apharamceutically acceptable excipient.

Also provided is a method of treating cancer in a subject in needthereof, the method comprising administering to the subject atherapeutically effective amount of a compound of the present invention,or a pharmaceutically acceptable salt thereof.

In some embodiments, a method is provided of treating a Rasprotein-related disorder in a subject in need thereof, the methodcomprising administering to the subject a therapeutically effectiveamount of a compound of the present invention, or a pharmaceuticallyacceptable salt thereof.

Further provided is a method of inhibiting a Ras protein in a cell, themethod comprising contacting the cell with an effective amount of acompound of the present invention, or a pharmaceutically acceptable saltthereof.

It is specifically contemplated that any limitation discussed withrespect to one embodiment of the invention may apply to any otherembodiment of the invention. Furthermore, any compound or composition ofthe invention may be used in any method of the invention, and any methodof the invention may be used to produce or to utilize any compound orcomposition of the invention.

Definitions and Chemical Terms

In this application, unless otherwise clear from context, (i) the term“a” means “one or more”; (ii) the term “or” is used to mean “and/or”unless explicitly indicated to refer to alternatives only or thealternative are mutually exclusive, although the disclosure supports adefinition that refers to only alternatives and “and/or”; (iii) theterms “comprising” and “including” are understood to encompass itemizedcomponents or steps whether presented by themselves or together with oneor more additional components or steps; and (iv) where ranges areprovided, endpoints are included.

As used herein, the term “about” is used to indicate that a valueincludes the standard deviation of error for the device or method beingemployed to determine the value. In certain embodiments, the term“about” refers to a range of values that fall within 25%, 20%, 19%, 18%,17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%,1%, or less in either direction (greater than or less than) of a statedvalue, unless otherwise stated or otherwise evident from the context(e.g., where such number would exceed 100% of a possible value).

As used herein, the term “adjacent” in the context of describingadjacent atoms refers to bivalent atoms that are directly connected by acovalent bond.

A “compound of the present invention” and similar terms as used herein,whether explicitly noted or not, refers to Ras inhibitors describedherein, including compounds of Formula Ia or Formula Ib and subformulaethereof, and compounds of Table 1a or Table 1b, as well as salts (e.g.,pharmaceutically acceptable salts), solvates, hydrates, stereoisomers(including atropisomers), and tautomers thereof.

The term “wild-type” refers to an entity having a structure or activityas found in nature in a “normal” (as contrasted with mutant, diseased,altered, etc.) state or context. Those of ordinary skill in the art willappreciate that wild-type genes and polypeptides often exist in multipledifferent forms (e.g., alleles).

Those skilled in the art will appreciate that certain compoundsdescribed herein can exist in one or more different isomeric (e.g.,stereoisomers, geometric isomers, atropisomers, tautomers) or isotopic(e.g., in which one or more atoms has been substituted with a differentisotope of the atom, such as hydrogen substituted for deuterium) forms.Unless otherwise indicated or clear from context, a depicted structurecan be understood to represent any such isomeric or isotopic form,individually or in combination.

Compounds described herein can be asymmetric (e.g., having one or morestereocenters). All stereoisomers, such as enantiomers anddiastereomers, are intended unless otherwise indicated. Compounds of thepresent disclosure that contain asymmetrically substituted carbon atomscan be isolated in optically active or racemic forms. Methods on how toprepare optically active forms from optically active starting materialsare known in the art, such as by resolution of racemic mixtures or bystereoselective synthesis. Many geometric isomers of olefins, C═N doublebonds, and the like can also be present in the compounds describedherein, and all such stable isomers are contemplated in the presentdisclosure. Cis and trans geometric isomers of the compounds of thepresent disclosure are described and may be isolated as a mixture ofisomers or as separated isomeric forms.

In some embodiments, one or more compounds depicted herein may exist indifferent tautomeric forms. As will be clear from context, unlessexplicitly excluded, references to such compounds encompass all suchtautomeric forms. In some embodiments, tautomeric forms result from theswapping of a single bond with an adjacent double bond and theconcomitant migration of a proton. In certain embodiments, a tautomericform may be a prototropic tautomer, which is an isomeric protonationstate having the same empirical formula and total charge as a referenceform. Examples of moieties with prototropic tautomeric forms areketone-enol pairs, amide-imidic acid pairs, lactam-lactim pairs,amide-imidic acid pairs, enamine-imine pairs, and annular forms where aproton can occupy two or more positions of a heterocyclic system, suchas, 1H- and 3H-imidazole, 1H-, 2H- and 4H-1,2,4-triazole, 1H- and2H-isoindole, and 1H- and 2H-pyrazole. In some embodiments, tautomericforms can be in equilibrium or sterically locked into one form byappropriate substitution. In certain embodiments, tautomeric formsresult from acetal interconversion.

Unless otherwise stated, structures depicted herein are also meant toinclude compounds that differ only in the presence of one or moreisotopically enriched atoms. Exemplary isotopes that can be incorporatedinto compounds of the present invention include isotopes of hydrogen,carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine, andiodine, such as ²H, ³H, ¹¹C, ¹³C, ¹⁴C, ¹³N, ¹⁵N, ¹⁵O, ¹⁷O, ¹⁸O, ³²P,³³P, ³⁵S, ¹⁸F, ³⁶Cl, ¹²³I and ¹²⁵I. Isotopically-labeled compounds(e.g., those labeled with ³H and ¹⁴C) can be useful in compound orsubstrate tissue distribution assays. Tritiated (i.e., ³H) and carbon-14(i.e., ¹⁴C) isotopes can be useful for their ease of preparation anddetectability. Further, substitution with heavier isotopes such asdeuterium (i.e., ²H) may afford certain therapeutic advantages resultingfrom greater metabolic stability (e.g., increased in vivo half-life orreduced dosage requirements). In some embodiments, one or more hydrogenatoms are replaced by ²H or ³H, or one or more carbon atoms are replacedby ¹³C- or ¹⁴C-enriched carbon. Positron emitting isotopes such as ¹⁵O,¹³N, ¹¹C, and ¹⁸F are useful for positron emission tomography (PET)studies to examine substrate receptor occupancy. Preparations ofisotopically labelled compounds are known to those of skill in the art.For example, isotopically labeled compounds can generally be prepared byfollowing procedures analogous to those disclosed for compounds of thepresent invention described herein, by substituting an isotopicallylabeled reagent for a non-isotopically labeled reagent.

As is known in the art, many chemical entities can adopt a variety ofdifferent solid forms such as, for example, amorphous forms orcrystalline forms (e.g., polymorphs, hydrates, solvate). In someembodiments, compounds of the present invention may be utilized in anysuch form, including in any solid form. In some embodiments, compoundsdescribed or depicted herein may be provided or utilized in hydrate orsolvate form.

At various places in the present specification, substituents ofcompounds of the present disclosure are disclosed in groups or inranges. It is specifically intended that the present disclosure includeeach and every individual subcombination of the members of such groupsand ranges. For example, the term “C₁-C₆ alkyl” is specifically intendedto individually disclose methyl, ethyl, C₃ alkyl, C₄ alkyl, C₅ alkyl,and C₆ alkyl. Furthermore, where a compound includes a plurality ofpositions at which substituents are disclosed in groups or in ranges,unless otherwise indicated, the present disclosure is intended to coverindividual compounds and groups of compounds (e.g., genera andsubgenera) containing each and every individual subcombination ofmembers at each position.

The term “optionally substituted X” (e.g., “optionally substitutedalkyl”) is intended to be equivalent to “X, wherein X is optionallysubstituted” (e.g., “alkyl, wherein said alkyl is optionallysubstituted”). It is not intended to mean that the feature “X” (e.g.,alkyl) per se is optional. As described herein, certain compounds ofinterest may contain one or more “optionally substituted” moieties. Ingeneral, the term “substituted”, whether preceded by the term“optionally” or not, means that one or more hydrogens of the designatedmoiety are replaced with a suitable substituent, e.g., any of thesubstituents or groups described herein. Unless otherwise indicated, an“optionally substituted” group may have a suitable substituent at eachsubstitutable position of the group, and when more than one position inany given structure may be substituted with more than one substituentselected from a specified group, the substituent may be either the sameor different at every position. For example, in the term “optionallysubstituted C₁-C₆ alkyl-C₂-C₉ heteroaryl,” the alkyl portion, theheteroaryl portion, or both, may be optionally substituted. Combinationsof substituents envisioned by the present disclosure are preferablythose that result in the formation of stable or chemically feasiblecompounds. The term “stable”, as used herein, refers to compounds thatare not substantially altered when subjected to conditions to allow fortheir production, detection, and, in certain embodiments, theirrecovery, purification, and use for one or more of the purposesdisclosed herein.

Suitable monovalent substituents on a substitutable carbon atom of an“optionally substituted” group may be, independently, deuterium;halogen; —(CH₂)₀₋₄R^(∘); —(CH₂)₀₋₄OR^(∘); —O(CH₂)₀₋₄R^(∘);—O—(CH₂)₀₋₄C(O)OR^(∘); —(CH₂)₀₋₄CH(OR^(∘))₂; —(CH₂)₀₋₄SR^(∘);—(CH₂)₀₋₄Ph, which may be substituted with R^(∘); —(CH₂)₀₋₄O(CH₂)₀₋₁Phwhich may be substituted with R^(∘); —CH═CHPh, which may be substitutedwith R^(∘); —(CH₂)₀₋₄O(CH₂)₀₋₁-pyridyl which may be substituted withR^(∘); 4-11 membered saturated or unsaturated heterocycloalkyl (e.g.,4-8 membered saturated or unsaturated heterocycloalkyl (e.g., pyridyl))which may be further optionally substituted (e.g., with a methyl); 3-8membered saturated or unsaturated cycloalkyl (e.g., cyclopropyl,cyclobutyl, or cyclopentyl); —NO₂; —CN; —N₃; —(CH₂)₀₋₄N(R^(∘))₂;—(CH₂)₀₋₄N(R^(∘))C(O)R^(∘); —N(R^(∘))C(S)R^(∘);—(CH₂)₀₋₄N(R^(∘))C(O)NR^(∘) ₂; —N(R^(∘))C(S)NR^(∘) ₂;—(CH₂)₀₋₄N(R^(∘))C(O)OR^(∘); —N(R^(∘))N(R^(∘))C(O)R^(∘);—N(R^(∘))N(R^(∘))C(O)NR^(∘) ₂; —N(R^(∘))N(R^(∘))C(O)OR^(∘);—(CH₂)₀₋₄C(O)R^(∘); —C(S)R^(∘); —(CH₂)₀₋₄C(O)OR^(∘);—(CH₂)₀₋₄—C(O)—N(R^(∘))₂; —(CH₂)₀₋₄—C(O)—N(R^(∘))—S(O)₂—R^(∘);—C(NCN)NR^(∘) ₂; —(CH₂)₀₋₄C(O)SR^(∘); —(CH₂)₀₋₄C(O)OSiR^(∘) ₃;—(CH₂)₀₋₄OC(O)R^(∘); —OC(O)(CH₂)₀₋₄SR^(∘); —SC(S)SR^(∘);—(CH₂)₀₋₄SC(O)R^(∘); —(CH₂)₀₋₄C(O)NR^(∘) ₂; —C(S)NR^(∘) ₂; —C(S)SR^(∘);—(CH₂)₀₋₄OC(O)NR^(∘) ₂; —C(O)N(OR^(∘))R^(∘); —C(O)C(O)R^(∘);—C(O)CH₂C(O)R^(∘); —C(NOR^(∘))R^(∘); —(CH₂)₀₋₄SSR^(∘);—(CH₂)₀₋₄S(O)₂R^(∘); —(CH₂)₀₋₄S(O)₂OR^(∘); —(CH₂)₀₋₄OS(O)₂R^(∘);—S(O)₂NR^(∘) ₂; —(CH₂)₀₋₄S(O)R^(∘); —N(R^(∘))S(O)₂NR^(∘) ₂;—N(R^(∘))S(O)₂R^(∘); —N(OR^(∘))R^(∘); —C(NOR^(∘))NR^(∘) ₂; —C(NH)NR^(∘)₂; —P(O)₂R^(∘); —P(O)R^(∘) ₂; —P(O)(OR^(∘))₂; —OP(O)R^(∘) ₂;—OP(O)(OR^(∘))₂; —OP(O)(OR^(∘))R^(∘), —SiR^(∘) ₃; —(C₁₋₄ straight orbranched alkylene)O—N(R^(∘))₂; or —(C₁₋₄ straight or branchedalkylene)C(O)O—N(R^(∘))₂, wherein each R^(∘) may be substituted asdefined below and is independently hydrogen, —C₁₋₆ aliphatic, —CH₂Ph,—O(CH₂)₀₋₁Ph, —CH₂-(5-6 membered heteroaryl ring), or a 3-6-memberedsaturated, partially unsaturated, or aryl ring having 0-4 heteroatomsindependently selected from nitrogen, oxygen, or sulfur, or,notwithstanding the definition above, two independent occurrences ofR^(∘), taken together with their intervening atom(s), form a3-12-membered saturated, partially unsaturated, or aryl mono- orbicyclic ring having 0-4 heteroatoms independently selected fromnitrogen, oxygen, or sulfur, which may be substituted as defined below.

Suitable monovalent substituents on R^(∘) (or the ring formed by takingtwo independent occurrences of R^(∘) together with their interveningatoms), may be, independently, halogen, —(CH₂)₀₋₂R^(●), -(haloR^(●)),—(CH₂)₀₋₂OH, —(CH₂)₀₋₂OR^(●), —(CH₂)₀₋₂CH(OR^(●))₂; —O(haloR^(●)), —CN,—N₃, —(C H₂)₀₋₂C(O)R^(●), —(CH₂)₀₋₂C(O)OH, —(CH₂)₀₋₂C(O)OR^(●),—(CH₂)₀₋₂SR^(●), —(CH₂)₀₋₂SH, —(CH₂)₀₋₂NH₂, —(CH₂)₀₋₂NH R^(●),—(CH₂)₀₋₂NR^(●) ₂, —NO₂, —SiR^(●) ₃, —OSiR^(●) ₃, —C(O)SR^(●)—(C₁₋₄straight or branched alkylene)C(O)OR^(●), or —SSR^(●) wherein each R^(●)is unsubstituted or where preceded by “halo” is substituted only withone or more halogens, and is independently selected from C₁₋₄ aliphatic,—CH₂Ph, —O(CH₂)₀₋₁Ph, or a 5-6-membered saturated, partiallyunsaturated, or aryl ring having 0-4 heteroatoms independently selectedfrom nitrogen, oxygen, or sulfur. Suitable divalent substituents on asaturated carbon atom of R^(∘) include ═O and ═S.

Suitable divalent substituents on a saturated carbon atom of an“optionally substituted” group include the following: ═O, ═S, ═NNR*₂,═NNHC(O)R*, ═NNHC(O)OR*, ═NNHS(O)₂R*, ═NR*, ═NOR*, —O(C(R*₂))₂₋₃O—, or—S(C(R*₂))₂₋₃S—, wherein each independent occurrence of R* is selectedfrom hydrogen, C₁₋₆ aliphatic which may be substituted as defined below,or an unsubstituted 5-6-membered saturated, partially unsaturated, oraryl ring having 0-4 heteroatoms independently selected from nitrogen,oxygen, or sulfur. Suitable divalent substituents that are bound tovicinal substitutable carbons of an “optionally substituted” groupinclude: —O(CR*₂)₂₋₃O—, wherein each independent occurrence of R* isselected from hydrogen, C₁₋₆ aliphatic which may be substituted asdefined below, or an unsubstituted 5-6-membered saturated, partiallyunsaturated, or aryl ring having 0-4 heteroatoms independently selectedfrom nitrogen, oxygen, or sulfur.

Suitable substituents on the aliphatic group of R* include halogen,—R^(●), -(haloR^(●)), —OH, —OR^(●), —O(haloR^(●)), —CN, —C(O)OH,—C(O)OR^(●), —NH₂, —NHR^(●), —NR^(●) ₂, or—NO₂, wherein each R^(●) isunsubstituted or where preceded by “halo” is substituted only with oneor more halogens, and is independently C₁₋₄ aliphatic, —CH₂Ph,—O(CH₂)₀₋₁Ph, or a 5-6-membered saturated, partially unsaturated, oraryl ring having 0-4 heteroatoms independently selected from nitrogen,oxygen, or sulfur.

Suitable substituents on a substitutable nitrogen of an “optionallysubstituted” group include —R^(†), —NR^(†) ₂, —C(O)R^(†), —C(O)OR^(†),—C(O)C(O)R^(†), —C(O)CH₂C(O)R^(†), —S(O)₂R^(†), —S(O)₂NR^(†) ₂,—C(S)NR^(†) ₂, —C(NH)NR^(†)2, or—N(R^(†))S(O)₂R^(†); wherein each R^(†)is independently hydrogen, C₁₋₆ aliphatic which may be substituted asdefined below, unsubstituted —OPh, or an unsubstituted 3-6-memberedsaturated, partially unsaturated, or aryl ring having 0-4 heteroatomsindependently selected from nitrogen, oxygen, or sulfur, or,notwithstanding the definition above, two independent occurrences ofR^(†), taken together with their intervening atom(s) form anunsubstituted 3-12-membered saturated, partially unsaturated, or arylmono- or bicyclic ring having 0-4 heteroatoms independently selectedfrom nitrogen, oxygen, or sulfur.

Suitable substituents on an aliphatic group of R^(†) are independentlyhalogen, —R^(●), -(haloR^(●)), —OH, —OR^(●), —O(haloR^(●)), —CN,—C(O)OH, —C(O)OR^(●), —NH₂, —NHR^(●), —NR^(●) ₂, or —NO₂, wherein eachR^(●) is unsubstituted or where preceded by “halo” is substituted onlywith one or more halogens, and is independently C₁₋₄ aliphatic, —CH₂Ph,—O(CH₂)₀₋₁Ph, or a 5-6-membered saturated, partially unsaturated, oraryl ring having 0-4 heteroatoms independently selected from nitrogen,oxygen, or sulfur. Suitable divalent substituents on a saturated carbonatom of R^(†) include ═O and ═S.

The term “acetyl,” as used herein, refers to the group —C(O)CH₃.

The term “alkoxy,” as used herein, refers to a —O—C₁-C₂₀ alkyl group,wherein the alkoxy group is attached to the remainder of the compoundthrough an oxygen atom.

The term “alkyl,” as used herein, refers to a saturated, straight orbranched monovalent hydrocarbon group containing from 1 to 20 (e.g.,from 1 to 10 or from 1 to 6) carbons. In some embodiments, an alkylgroup is unbranched (i.e., is linear); in some embodiments, an alkylgroup is branched. Alkyl groups are exemplified by, but not limited to,methyl, ethyl, n- and iso-propyl, n-, sec-, iso- and tert-butyl, andneopentyl.

The term “alkylene,” as used herein, represents a saturated divalenthydrocarbon group derived from a straight or branched chain saturatedhydrocarbon by the removal of two hydrogen atoms, and is exemplified bymethylene, ethylene, isopropylene, and the like. The term “C_(x)-C_(y)alkylene” represents alkylene groups having between x and y carbons.Exemplary values for x are 1, 2, 3, 4, 5, and 6, and exemplary valuesfor y are 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 16, 18, or 20 (e.g.,C₁-C₆, C₁-C₁₀, C₂-C₂₀, C₂-C₆, C₂-C₁₀, or C₂-C₂₀ alkylene). In someembodiments, the alkylene can be further substituted with 1, 2, 3, or 4substituent groups as defined herein.

The term “alkenyl,” as used herein, represents monovalent straight orbranched chain groups of, unless otherwise specified, from 2 to 20carbons (e.g., from 2 to 6 or from 2 to 10 carbons) containing one ormore carbon-carbon double bonds and is exemplified by ethenyl,1-propenyl, 2-propenyl, 2-methyl-1-propenyl, 1-butenyl, and 2-butenyl.Alkenyls include both cis and trans isomers. The term “alkenylene,” asused herein, represents a divalent straight or branched chain groups of,unless otherwise specified, from 2 to 20 carbons (e.g., from 2 to 6 orfrom 2 to 10 carbons) containing one or more carbon-carbon double bonds.

The term “alkynyl,” as used herein, represents monovalent straight orbranched chain groups from 2 to 20 carbon atoms (e.g., from 2 to 4, from2 to 6, or from 2 to 10 carbons) containing a carbon-carbon triple bondand is exemplified by ethynyl, and 1-propynyl.

The term “alkynyl sulfone,” as used herein, represents a groupcomprising the structure

wherein R is any chemically feasible substituent described herein.

The term “amino,” as used herein, represents —N(R^(†))₂, e.g., —NH₂ and—N(CH₃)₂.

The term “aminoalkyl,” as used herein, represents an alkyl moietysubstituted on one or more carbon atoms with one or more amino moieties.

The term “amino acid,” as described herein, refers to a molecule havinga side chain, an amino group, and an acid group (e.g., —CO₂H or—SO₃H),wherein the amino acid is attached to the parent molecular group by theside chain, amino group, or acid group (e.g., the side chain). As usedherein, the term “amino acid” in its broadest sense, refers to anycompound or substance that can be incorporated into a polypeptide chain,e.g., through formation of one or more peptide bonds. In someembodiments, an amino acid has the general structure H₂N—C(H)(R)—COOH.In some embodiments, an amino acid is a naturally-occurring amino acid.In some embodiments, an amino acid is a synthetic amino acid; in someembodiments, an amino acid is a D-amino acid; in some embodiments, anamino acid is an L-amino acid. “Standard amino acid” refers to any ofthe twenty standard L-amino acids commonly found in naturally occurringpeptides. Exemplary amino acids include alanine, arginine, asparagine,aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine,optionally substituted hydroxylnorvaline, isoleucine, leucine, lysine,methionine, norvaline, ornithine, phenylalanine, proline, pyrrolysine,selenocysteine, serine, taurine, threonine, tryptophan, tyrosine, andvaline.

The term “aryl,” as used herein, represents a monovalent monocyclic,bicyclic, or multicyclic ring system formed by carbon atoms, wherein thering attached to the pendant group is aromatic. Examples of aryl groupsare phenyl, naphthyl, phenanthrenyl, and anthracenyl. An aryl ring canbe attached to its pendant group at any heteroatom or carbon ring atomthat results in a stable structure and any of the ring atoms can beoptionally substituted unless otherwise specified.

The term “C₀,” as used herein, represents a bond. For example, part ofthe term —N(C(O)—(C₀-C₅ alkylene-H)— includes —N(C(O)—(C₀ alkylene-H)—,which is also represented by —N(C(O)—H)—.

The terms “carbocyclic” and “carbocyclyl,” as used herein, refer to amonovalent, optionally substituted C₃-C₁₂ monocyclic, bicyclic, ortricyclic ring structure, which may be bridged, fused or spirocyclic, inwhich all the rings are formed by carbon atoms and at least one ring isnon-aromatic. Carbocyclic structures include cycloalkyl, cycloalkenyl,and cycloalkynyl groups. Examples of carbocyclyl groups are cyclohexyl,cyclohexenyl, cyclooctynyl, 1,2-dihydronaphthyl,1,2,3,4-tetrahydronaphthyl, fluorenyl, indenyl, indanyl, decalinyl, andthe like. A carbocyclic ring can be attached to its pendant group at anyring atom that results in a stable structure and any of the ring atomscan be optionally substituted unless otherwise specified.

The term “carbonyl,” as used herein, represents a C(O) group, which canalso be represented as C═O.

The term “carboxyl,” as used herein, means —CO₂H, (C═O)(OH), COOH, orC(O)OH or the unprotonated counterparts.

The term “cyano,” as used herein, represents a —CN group.

The term “cycloalkyl,” as used herein, represents a monovalent saturatedcyclic hydrocarbon group, which may be bridged, fused or spirocyclichaving from three to eight ring carbons, unless otherwise specified, andis exemplified by cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,cycloheptyl, and cycloheptyl.

The term “cycloalkenyl,” as used herein, represents a monovalent,non-aromatic, saturated cyclic hydrocarbon group, which may be bridged,fused or spirocyclic having from three to eight ring carbons, unlessotherwise specified, and containing one or more carbon-carbon doublebonds.

The term “diastereomer,” as used herein, means stereoisomers that arenot mirror images of one another and are non-superimposable on oneanother.

The term “enantiomer,” as used herein, means each individual opticallyactive form of a compound of the invention, having an optical purity orenantiomeric excess (as determined by methods standard in the art) of atleast 80% (i.e., at least 90% of one enantiomer and at most 10% of theother enantiomer), preferably at least 90% and more preferably at least98%.

The term “haloacetyl,” as used herein, refers to an acetyl group whereinat least one of the hydrogens has been replaced by a halogen.

The term “haloalkyl,” as used herein, represents an alkyl moietysubstituted on one or more carbon atoms with one or more of the same ofdifferent halogen moieties.

The term “halogen,” as used herein, represents a halogen selected frombromine, chlorine, iodine, or fluorine.

The term “heteroalkyl,” as used herein, refers to an “alkyl” group, asdefined herein, in which at least one carbon atom has been replaced witha heteroatom (e.g., an O, N, or S atom). The heteroatom may appear inthe middle or at the end of the radical.

The term “heteroaryl,” as used herein, represents a monovalent,monocyclic or polycyclic ring structure that contains at least one fullyaromatic ring: i.e., they contain 4n+2 pi electrons within themonocyclic or polycyclic ring system and contains at least one ringheteroatom selected from N, O, or S in that aromatic ring. Exemplaryunsubstituted heteroaryl groups are of 1 to 12 (e.g., 1 to 11, 1 to 10,1 to 9, 2 to 12, 2 to 11, 2 to 10, or 2 to 9) carbons. The term“heteroaryl” includes bicyclic, tricyclic, and tetracyclic groups inwhich any of the above heteroaromatic rings is fused to one or more,aryl or carbocyclic rings, e.g., a phenyl ring, or a cyclohexane ring.Examples of heteroaryl groups include, but are not limited to, pyridyl,pyrazolyl, benzooxazolyl, benzoimidazolyl, benzothiazolyl, imidazolyl,thiazolyl, quinolinyl, tetrahydroquinolinyl, and 4-azaindolyl. Aheteroaryl ring can be attached to its pendant group at any ring atomthat results in a stable structure and any of the ring atoms can beoptionally substituted unless otherwise specified. In some embodiment,the heteroaryl is substituted with 1, 2, 3, or 4 substituents groups.

The term “heterocycloalkyl,” as used herein, represents a monovalentmonocyclic, bicyclic or polycyclic ring system, which may be bridged,fused or spirocyclic, wherein at least one ring is non-aromatic andwherein the non-aromatic ring contains one, two, three, or fourheteroatoms independently selected from the group consisting ofnitrogen, oxygen, and sulfur. The 5-membered ring has zero to two doublebonds, and the 6- and 7-membered rings have zero to three double bonds.Exemplary unsubstituted heterocycloalkyl groups are of 1 to 12 (e.g., 1to 11, 1 to 10, 1 to 9, 2 to 12, 2 to 11, 2 to 10, or 2 to 9) carbons.The term “heterocycloalkyl” also represents a heterocyclic compoundhaving a bridged multicyclic structure in which one or more carbons orheteroatoms bridges two non-adjacent members of a monocyclic ring, e.g.,a quinuclidinyl group. The term “heterocycloalkyl” includes bicyclic,tricyclic, and tetracyclic groups in which any of the above heterocyclicrings is fused to one or more aromatic, carbocyclic, heteroaromatic, orheterocyclic rings, e.g., an aryl ring, a cyclohexane ring, acyclohexene ring, a cyclopentane ring, a cyclopentene ring, a pyridinering, or a pyrrolidine ring. Examples of heterocycloalkyl groups arepyrrolidinyl, piperidinyl, 1,2,3,4-tetrahydroquinolinyl,decahydroquinolinyl, dihydropyrrolopyridine, and decahydronapthyridinyl.A heterocycloalkyl ring can be attached to its pendant group at any ringatom that results in a stable structure and any of the ring atoms can beoptionally substituted unless otherwise specified.

The term “hydroxy,” as used herein, represents a —OH group.

The term “hydroxyalkyl,” as used herein, represents an alkyl moietysubstituted on one or more carbon atoms with one or more —OH moieties.

The term “isomer,” as used herein, means any tautomer, stereoisomer,atropiosmer, enantiomer, or diastereomer of any compound of theinvention. It is recognized that the compounds of the invention can haveone or more chiral centers or double bonds and, therefore, exist asstereoisomers, such as double-bond isomers (i.e., geometric E/Z isomers)or diastereomers (e.g., enantiomers (i.e., (+) or (−)) or cis/transisomers). According to the invention, the chemical structures depictedherein, and therefore the compounds of the invention, encompass all thecorresponding stereoisomers, that is, both the stereomerically pure form(e.g., geometrically pure, enantiomerically pure, or diastereomericallypure) and enantiomeric and stereoisomeric mixtures, e.g., racemates.Enantiomeric and stereoisomeric mixtures of compounds of the inventioncan typically be resolved into their component enantiomers orstereoisomers by well-known methods, such as chiral-phase gaschromatography, chiral-phase high performance liquid chromatography,crystallizing the compound as a chiral salt complex, or crystallizingthe compound in a chiral solvent. Enantiomers and stereoisomers can alsobe obtained from stereomerically or enantiomerically pure intermediates,reagents, and catalysts by well-known asymmetric synthetic methods.

The term “stereoisomer,” as used herein, refers to all possibledifferent isomeric as well as conformational forms which a compound maypossess (e.g., a compound of any formula described herein), inparticular all possible stereochemically and conformationally isomericforms, all diastereomers, enantiomers or conformers of the basicmolecular structure, including atropisomers. Some compounds of thepresent invention may exist in different tautomeric forms, all of thelatter being included within the scope of the present invention.

The term “sulfonyl,” as used herein, represents an —S(O)₂— group.

The term “thiocarbonyl,” as used herein, refers to a —C(S)— group.

Those of ordinary skill in the art, reading the present disclosure, willappreciate that certain compounds described herein may be provided orutilized in any of a variety of forms such as, for example, salt forms,protected forms, pro-drug forms, ester forms, isomeric forms (e.g.,optical or structural isomers), isotopic forms, etc. In someembodiments, reference to a particular compound may relate to a specificform of that compound. In some embodiments, reference to a particularcompound may relate to that compound in any form. In some embodiments,for example, a preparation of a single stereoisomer of a compound may beconsidered to be a different form of the compound than a racemic mixtureof the compound; a particular salt of a compound may be considered to bea different form from another salt form of the compound; a preparationcontaining one conformational isomer ((Z) or (E)) of a double bond maybe considered to be a different form from one containing the otherconformational isomer ((E) or (Z)) of the double bond; a preparation inwhich one or more atoms is a different isotope than is present in areference preparation may be considered to be a different form.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1A is a graph demonstrating the in vivo efficacy of Compound A, aRas inhibitor disclosed herein, in a human pancreatic adenocarcinomaHPAC KRAS^(G12D/wt) xenograft model using female BALB/c nude mice. Thegraph shows tumor volume (mm³) vs. days post-implant of the mousexenograft model. Mice were randomized to treatment groups prior toadministration of test articles or vehicle. Compound A was administeredby oral gavage once every other day.

FIG. 1B is a graph demonstrating dose-dependent exposure of Compound A,a Ras inhibitor disclosed herein, in blood and tumor samples from BALB/cnude mice (6-8 weeks old, human non-small cell lung cancer (NSCLC)NCI-H441 KRAS^(G12V/wt) xenograft model), monitored through 72 hoursfollowing dose. Pharmacokinetics were analyzed based on totalconcentration (nM) of Compound A in tumors or blood, following a singleoral gavage dose of Compound A at 10, 25 or 50 mg/kg to 72 hours. Ateach time point, tumor or blood was sampled from n=3 animals.

FIG. 1C is a graph showing PK (10 mg/kg, po) and PD (% tumor DUSPrelative to control, 10 and 25 mg/kg, po) in naïve animals treated witha single dose of Compound A, a Ras inhibitor disclosed herein.

FIG. 1D is a graph demonstrating the in vivo efficacy of Compound A, aRas inhibitor disclosed herein, in the NCI-H441 CDX model withheterozygous KRAS^(G12V). NCI-H441 cells were implanted in 50% Matrigel.Animals were randomized and treatment was initiated at average tumorvolume of ˜155 mm³. Animals were dosed with Compound A 10 or 25 mg/kg poqd or Control for 32 days. All dose levels were tolerated, n=10/group.***p<0.0001 by one-way ANOVA.

FIG. 1E shows end of study responses for Compound A, a KRAS(ON)inhibitor disclosed herein, in the NCI-H441 CDX model with heterozygousKRAS^(G12V). NCI-H441 end of study tumors were graphed as % change intumor volume compared to volume at treatment initiation. R(regressions)=number of regressions >10% from initial. CR (completeresponse)=number of regressions >80% from initial. Each animalrepresented as a separate bar.

FIG. 1F shows shows % body weight change in animals treated withCompound A, a KRAS(ON) inhibitor disclosed herein, (in the NCI-H441 CDXmodel with heterozygous KRAS^(G12V)). NCI-H441 cell-derived xenograftswere measured twice weekly by caliper measurements. Body weight changegraphed as percentage of animals starting body weight.

FIG. 2A is a graph demonstrating in vivo efficacy of Compound A, aKRAS(ON) inhibitor disclosed herein, in the human pancreatic Capan-2 CDXxenograft model with heterozygous KRAS^(G12V) using female BALB/c nudemice. The graph shows tumor volume (mm³) vs. days post-implant of themouse xenograft model. Capan-2 cells were implanted in 50% Matrigel.Animals were randomized and treatment was initiated at average tumorvolume of ˜166 mm³. Animals were dosed with Compound A 10 mg/kg po qd or25 mg/kg po q2d or Control for 28 days. All dose levels were tolerated,n=8/group. **p=0.01, ***p<0.0001 by one-way ANOVA.

FIG. 2B shows end of study responses for Compound A, a KRAS(ON)inhibitor disclosed herein, in the human pancreatic Capan-2 CDXxenograft model with heterozygous KRAS^(G12V). Capan-2 end of studytumors were graphed as % change in tumor volume compared to volume attreatment initiation. R (regressions)=number of regressions >10% frominitial. Each animal represented as a separate bar.

FIG. 2C shows % body weight change in animals treated with Compound A, aKRAS(ON) inhibitor disclosed herein, in the human pancreatic Capan-2 CDXxenograft model with heterozygous KRAS^(G12V). Capan-2 cell-derivedxenografts were measured twice weekly by caliper measurements. Bodyweight change graphed as percentage of animals starting body weight.

FIG. 2D is a graph demonstrating in vivo efficacy of Compound A, aKRAS(ON) inhibitor disclosed herein, in the human colorectal SW403KRAS^(G12V/wt) xenograft model using female BALB/c nude mice. The graphshows tumor volume (mm³) vs. days post-implant of the mouse xenograftmodel. SW403 cells were implanted in 50% Matrigel. Animals wererandomized and treatment was initiated at average tumor volume of ˜171mm³. Animals were dosed with Compound A 25 mg/kg po qd or 50 mg/kg poq2d or Control for 28 days. All dose levels were tolerated, n=8/group.***p<0.0001 by one-way ANOVA.

FIG. 2E shows end of study responses for Compound A, a KRAS(ON)inhibitor disclosed herein, in the human colorectal SW403 KRAS^(G12V/wt)xenograft model. SW403 end of study tumors were graphed as % change intumor volume compared to volume at treatment initiation. R(regressions)=number of regressions >10% from initial. CR (completeresponse)=number of regressions >80% from initial. Each animalrepresented as a separate bar.

FIG. 2F shows % body weight change in animals treated with Compound A, aKRAS(ON) inhibitor disclosed herein, in the human colorectal SW403KRAS^(G12V/wt) xenograft model. SW403 cell-derived xenografts weremeasured twice weekly by caliper measurements. Body weight changegraphed as percentage of animals starting body weight.

FIG. 3A demonstrates in vitro efficacy of Compound A, a KRAS(ON)inhibitor disclosed herein, in multiple RAS-driven cancer cell lines.Each graph shows cell proliferation (% relative to control) vs. log M[Compound A], Potency of in vitro cell proliferation inhibition ofCapan-1 (KRAS^(G12V)), NCI-H358 (KRAS^(G12C)), AsPC-1 (KRAS^(G12D)),HCT116 (KRAS^(G13D)), SK-MEL-30 (NRAS^(Q61K)), NCI-H1975(EGFR^(T790M/L858R)), and A375 (BRAF^(V600E)) cells exposed to CompoundA for 120 hours.

FIG. 3B demonstrates in vitro efficacy of Compound B, a KRAS(ON)inhibitor disclosed herein, in multiple RAS-driven cancer cell lines.Each graph shows cell proliferation (% relative to control) vs. log M[Compound B], Potency of in vitro cell proliferation inhibition ofCapan-1 (KRAS^(G12V)), NCI-H358 (KRAS^(G12C)), AsPC-1 (KRAS^(G12D)),HCT116 (KRAS^(G13D)), SK-MEL-30 (NRAS^(Q61K)), NCI-H1975(EGFR^(T790M/L858R)), and A375 (BRAF^(V600E)) cells exposed to CompoundB for 120 hours.

FIG. 3C demonstrates in vitro efficacy of Compound C, a KRAS(ON)inhibitor disclosed herein, in multiple RAS-driven cancer cell lines.Each graph shows cell proliferation (% relative to control) vs. log M[Compound C], Potency of in vitro cell proliferation inhibition ofCapan-1 (KRAS^(G12V)), NCI-H358 (KRAS^(G12C)), AsPC-1 (KRAS^(G12D)), andA375 (BRAF^(V600E)) cells exposed to Compound C for 120 hours.

FIG. 4A demonstrates in vivo efficacy of Compound A (25 mg/kg po qd), aKRAS(ON) inhibitor disclosed herein, in the human pancreaticadenocarcinoma HPAC KRAS^(G12D/wt) xenograft model using female BALB/cnude mice. The graph shows tumor volume (mm³) vs. days post-implant ofthe mouse xenograft model. HPAC cells were implanted in 50% Matrigel.Animals were randomized and treatment was initiated at average tumorvolume of ˜142 mm³. Animals were dosed with Compound A 25 mg/kg po qd orControl for 28 days. Dose level was tolerated, n=9-10/group. ***p<0.0001by one-way ANOVA.

FIG. 4B shows end of study responses for Compound A, a KRAS(ON)inhibitor disclosed herein, in the human pancreatic adenocarcinoma HPACKRAS^(G12D/wt) xenograft model. HPAC end of study tumors were graphed as% change in tumor volume compared to volume at treatment initiation. CR(complete response)=number of regressions >80% from initial. Each animalrepresented as a separate bar.

FIG. 4C shows % body weight change in animals treated with Compound A, aKRAS(ON) inhibitor disclosed herein, in the human pancreaticadenocarcinoma HPAC KRAS^(G12D/wt) xenograft model. HPAC cell-derivedxenografts were measured twice weekly by caliper measurements. Bodyweight change graphed as percentage of animals starting body weight.

FIG. 4D demonstrates in vivo efficacy of Compound A, a KRAS(ON)inhibitor disclosed herein, in the human colorectal GP2d KRAS^(G12D/wt)xenograft model using female BALB/c nude mice. The graph shows tumorvolume (mm³) vs. days post-implant of the mouse xenograft model. GP2dcells were implanted in 50% Matrigel. Animals were randomized andtreatment was initiated at average tumor volume of ˜154 mm³. Animalswere dosed with Compound A 25 mg/kg po qd or Control for 28 days. Doselevel was tolerated, n=10/group. ***p<0.0001 by one-way ANOVA.

FIG. 4E shows end of study responses for Compound A, a KRAS(ON)inhibitor disclosed herein, in the human colorectal GP2d KRAS^(G12D/wt)xenograft model. GP2d end of study tumors were graphed as % change intumor volume compared to volume at treatment initiation. Each animalrepresented as a separate bar.

FIG. 4F shows % body weight change in animals treated with Compound A, aKRAS(ON) inhibitor disclosed herein, in the human colorectal GP2dKRAS^(G12D/wt) xenograft model. GP2d cell-derived xenografts weremeasured twice weekly by caliper measurements. Body weight changegraphed as percentage of animals starting body weight.

FIG. 5A demonstrates in vitro efficacy of Compound A, a KRAS(ON)inhibitor disclosed herein, in down-regulating immune checkpointproteins in NCI-H358 KRAS^(G12C) Cells in Vitro. FIG. 5A shows cellsurface expression of PD-L1, PVR and CD73 on H358 cells following48-hour treatment with Compound A in the presence of Interferon gamma(IFNγ), as measured by flow cytometry. Each graph shows meanfluorescence intensity ((MFI), for each respective immune checkpointprotein) vs. log M [Compound A],

FIG. 5B demonstrates in vitro efficacy of Compound A, a KRAS(ON)inhibitor disclosed herein, in down-regulating immune checkpointproteins in SW900 KRAS^(G12C) Cells in Vitro. FIG. 5B shows cell surfaceexpression of PD-L1, PVR and CD73 on SW900 cells following 48-hourtreatment with Compound A in the presence of Interferon gamma (IFNγ), asmeasured by flow cytometry. Each graph shows mean fluorescence intensity((MFI), for each respective immune checkpoint protein) vs. log M[Compound A],

FIG. 5C demonstrates in vitro efficacy of Compound A, a KRAS(ON)inhibitor disclosed herein, in down-regulating immune checkpointproteins in Capan-2 KRAS^(G12C) Cells in Vitro. FIG. 5C shows cellsurface expression of PD-L1, PVR and CD73 on Capan-2 cells following48-hour treatment with Compound A in the presence of Interferon gamma(IFNγ), as measured by flow cytometry. Each graph shows meanfluorescence intensity ((MFI), for each respective immune checkpointprotein) vs. log M [Compound A].

FIGS. 6A-6B demonstrate that Compound A, a KRAS(ON) inhibitor disclosedherein, is active against RAS oncogene switching mutations observed inKRAS^(G12C)(OFF) resistance. FIG. 6A is a heatmap representing cellularRAS/RAF disruption assay results regarding various KRAS mutations in thepresence of different RAS inhibitors. FIG. 6B shows the IC₅₀ valueassociated with each colored bar of the heatmap.

FIGS. 7A-7D demonstrate that Compound A, a KRAS(ON) inhibitor disclosedherein, drives regressions of a syngeneic KRAS^(G12C) tumor model invivo and synergizes with anti-PD-1. eCT26 (CRC, KRAS^(G12C/G12C)ABCB1^(−/−)) I20 tumor growth in individual mice treated with: vehicleand Isotype (FIG. 7A); anti-PD-1 (FIG. 7B); Compound A (FIG. 7C); andCompound A+anti-PD-1 (FIG. 7D).

FIG. 8 demonstrates that Compound A, a KRAS(ON) inhibitor disclosedherein, in combination with anti-PD-1 is well tolerated in vivo in theeCT26 (CRC, KRAS^(G12C/G12C) ABCB1^(−/−) I20 model. Body weight changegraphed as percentage of animals starting body weight.

FIGS. 9A, 9B and 9C demonstrates that Compound A, a KRAS(ON) inhibitordisclosed herein, modulates the immune tumor microenvironment in favorof anti-tumor immunity in vivo. Flow cytometric immunophenotyping ofeCT26 (CRC, KRAS^(G12C/G12C) ABCB1^(−/−)) I20 tumors treated for 4 dayswith vehicle or Compound A 25 mg/kg qd. Symbols represent individualtumors. Average starting tumor volume was ˜188 mm³ for the vehicle groupand ˜586 mm³ for Compound A treated group. Single-agent Compound Aresulted in an increase of CD8+ T-cells (FIG. 9A). Compound A also ledto a decrease in M2 macrophages (FIG. 9B) and monocytic MDSCs (FIG. 9C).Data are mean+/−SD; *p<0.05, **p<0.01 by two-sided Student's t-test.

FIGS. 10A and 10B demonstrate that Compound A, a KRAS(ON) inhibitordisclosed herein, exhibits significant anti-tumor activity inKRAS^(G12X) tumor models of human NSCLC (FIG. 10A) or PDAC (FIG. 10B) invivo.

FIG. 11 demonstrates that Compound A, a KRAS(ON) inhibitor disclosedherein, extends time to tumor doubling across xenograft models. p<0.0001by Log-rank test (control vs treated). KRAS^(G12X): n=154; other RAS andRAS pathway mutations: n=86; All RAS Pathway^(MUT) includes both groups:n=240. Progression defined as tumor doubling from baseline over 28 days.

FIGS. 12A, 12B and 12C demonstrate that Compounds A, B and D, KRAS(ON)inhibitors disclosed herein, drive regressions of KRAS^(G12V) tumors invivo, as measured by mean tumor volume (FIG. 12A), % body weight change(FIG. 12B), and % change in tumor volume (FIG. 12C). n=6/group.***p<0.001. All treatments well tolerated as assessed by body weight.

FIGS. 13A and 13B show PD (FIG. 13A) and PK (FIG. 13B) resultsdemonstrating that Compounds A, B and D, KRAS(ON) inhibitors disclosedherein, deeply and durably inhibit RAS pathway signaling in vivo. Singledose experiment; all doses well tolerated.

DETAILED DESCRIPTION Compounds

Provided herein are Ras inhibitors. These Ras inhibitors target, thatis, selectively bind to or inhibit, Ras(ON) (e.g., selective over theGDP-bound, inactive state of Ras). As used herein, the term “RAS(ON)inhibitor” refers to an inhibitor that targets, that is, selectivelybinds to or inhibits, the GTP-bound, active state of RAS (e.g.,selective over the GDP-bound, inactive state of RAS). Inhibition of theGTP-bound, active state of RAS includes, for example, the inhibition ofoncogenic signaling from the GTP-bound, active state of RAS. In someembodiments, the RAS(ON) inhibitor is an inhibitor that selectivelybinds to and inhibits the GTP-bound, active state of RAS. In certainembodiments, RAS(ON) inhibitors may also bind to or inhibit theGDP-bound, inactive state of RAS (e.g., with a lower affinity orinhibition constant than for the GTP-bound, active state of RAS). Insome embodiments, a RAS(ON) inhibitor has a molecular weight of between800 and 1100 Da, inclusive. Accordingly, for example, the term “KRAS(ON)inhibitor” refers to any inhibitor that binds to KRAS in its GTP-bound“ON” position. A “KRAS^(G12C)(ON) inhibitor” is a KRAS inhibitor thatselectively binds to or targets the G12C mutant form of KRAS.Non-limiting examples of RAS(ON) inhibitors, some of which areKRAS^(G12C)(ON) inhibitors, are provided in WO 2021091982, WO2021091967, WO 2021091956, and WO 2020132597.

As used herein, the term “RAS(OFF) inhibitor” refers to an inhibitorthat targets, that is, selectively binds to or inhibits the GDP-bound,inactive state of RAS (e.g., selective over the GTP-bound, active stateof RAS). Inhibition of the GDP-bound, inactive state of RAS includes,for example, sequestering the inactive state by inhibiting the exchangeof GDP for GTP, thereby inhibiting RAS from adopting the activeconformation. In certain embodiments, RAS(OFF) inhibitors may also bindto or inhibit the GTP-bound, active state of RAS (e.g., with a loweraffinity or inhibition constant than for the GDP-bound, inactive stateof RAS). In some embodiments, a RAS(OFF) inhibitor has a molecularweight of under 700 Da. In some embodiments, a RAS(OFF) inhibitor has amolecular weight of under 700 Da. Accordingly, for example, the term“KRAS(OFF) inhibitor” refers to any inhibitor that binds to KRAS in itsGDP-bound “OFF” position. A “KRAS^(G12C)(OFF) inhibitor” is a KRASinhibitor that selectively binds to or targets the G12C mutant form ofKRAS. KRAS^(G12C)(OFF) inhibitors are known in the art and non-limitingexamples include adagrasib and sotorasib. Additional KRAS(OFF)inhibitors are provided herein.

The term “inhibitor” means a compound or agent (e.g., peptide, antibody)that prevents a biomolecule, (e.g., a protein) from completing orinitiating a reaction. An inhibitor can inhibit a reaction bycompetitive, uncompetitive, or non-competitive means.

The approach described herein entails formation of a high affinitythree-component complex between a synthetic ligand and two intracellularproteins which do not interact under normal physiological conditions:the target protein of interest (e.g., Ras), and a widely expressedcytosolic chaperone (presenter protein) in the cell (e.g., cyclophilinA). More specifically, in some embodiments, the inhibitors of Rasdescribed herein induce a new binding pocket in Ras by driving formationof a high affinity tri-complex between the Ras protein and the widelyexpressed cytosolic chaperone, cyclophilin A (CYPA). Without being boundby theory, the inventors believe that one way the inhibitory effect onRas is effected by compounds of the invention and the complexes theyform is by steric occlusion of the interaction site between Ras anddownstream effector molecules, such as RAF, which are required forpropagating the oncogenic signal.

Without being bound by theory, the inventors postulate that non-covalentinteractions of a compound of the present invention with Ras and thechaperone protein (e.g., cyclophilin A) may contribute to the inhibitionof Ras activity. For example, van der Waals, hydrophobic, hydrophilicand hydrogen bond interactions, and combinations thereof, may contributeto the ability of the compounds of the present invention to formcomplexes and act as Ras inhibitors. Accordingly, a variety of Rasproteins may be inhibited by compounds of the present invention (e.g., awild-type Ras or Ras^(amp), or K-Ras, N-Ras, H-Ras, and mutants thereofat positions 12, 13 and 61, such as G12C, G12D, G12V, G12S, G13C, G13D,and Q61L, and others described herein, as well as combinations of Rasproteins).

Accordingly, provided herein are compounds, or pharmaceuticallyacceptable salts thereof, having the structure of Formula Ia:

wherein A is optionally substituted 3 to 6-membered cycloalkylene,optionally substituted 3 to 6-membered heterocycloalkylene, optionallysubstituted 6-membered arylene, optionally substituted 5 to 6-memberedheteroarylene, optionally substituted C₂-C₄ alkylene, or optionallysubstituted C₂-C₄ alkenylene;

Y is

W is hydrogen, C₁-C₄ alkyl, optionally substituted C₁-C₃ heteroalkyl,optionally substituted 3 to 10-membered heterocycloalkyl, optionallysubstituted 3 to 10-membered cycloalkyl, optionally substituted 6 to10-membered aryl, or optionally substituted 5 to 10-membered heteroaryl;

X¹ and X⁴ are each, independently, CH₂ or NH;

R¹ is optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 15-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl; and

R² is hydrogen, optionally substituted C₁-C₆ alkyl, optionallysubstituted C₂-C₆ alkenyl, optionally substituted C₂-C₆ alkynyl,optionally substituted 3 to 6-membered cycloalkyl, optionallysubstituted 3 to 7-membered heterocycloalkyl, optionally substituted6-membered aryl, optionally substituted 5 or 6-membered heteroaryl; andR¹⁰ is hydrogen, hydroxy, optionally substituted C₁-C₃ alkyl, oroptionally substituted C₁-C₆ heteroalkyl.

In some embodiments, the compound, or pharmaceutically acceptable saltthereof, has the structure of Formula Ia-2:

wherein A is optionally substituted 3 to 6-membered cycloalkylene,optionally substituted 3 to 6-membered heterocycloalkylene, optionallysubstituted 6-membered arylene, or optionally substituted 5 to6-membered heteroarylene;

Y is

W is hydrogen, C₁-C₄ alkyl, optionally substituted 3 to 10-memberedheterocycloalkyl, optionally substituted 3 to 10-membered cycloalkyl,optionally substituted 6 to 10-membered aryl, or optionally substituted5 to 10-membered heteroaryl;

R¹ is optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 6-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl;

R² is hydrogen, optionally substituted C₁-C₆ alkyl, optionallysubstituted C₂-C₆ alkenyl, optionally substituted C₂-C₆ alkynyl,optionally substituted 3 to 6-membered cycloalkyl, optionallysubstituted 3 to 7-membered heterocycloalkyl, optionally substituted6-membered aryl, optionally substituted 5 or 6-membered heteroaryl; and

R¹⁰ is hydrogen or optionally substituted C₁-C₆ heteroalkyl. In someembodiments, R¹⁰ is hydrogen.

In some embodiments, R¹ is optionally substituted 6 to 10-membered arylor optionally substituted 5 to 10-membered heteroaryl. In someembodiments, R¹ is optionally substituted phenyl or optionallysubstituted pyridine.

In some embodiments, A is optionally substituted thiazole, optionallysubstituted triazole, optionally substituted morpholino, optionallysubstituted piperidinyl, optionally substituted pyridine, or optionallysubstituted phenyl. In some embodiments, A is optionally substitutedthiazole, optionally substituted triazole, optionally substitutedmorpholino, or phenyl. In some embodiments, A is not an optionallysubstituted phenyl or benzimidazole. In some embodiments, A is nothydroxyphenyl.

In some embodiments, Y is —NHC(O)— or —NHC(O)NH—.

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula IIa:

wherein a is 0 or 1.

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula IIa-1:

wherein X² is N or CH;

each R³ is independently selected from halogen, cyano, hydroxy,optionally substituted amine, optionally substituted amido, optionallysubstituted C₁-C₆ alkyl, optionally substituted C₁-C₆ heteroalkyl,optionally substituted 3 to 6-membered cycloalkyl, optionallysubstituted 3 to 6-membered cycloalkenyl, optionally substituted 3 to11-membered heterocycloalkyl (e.g., optionally substituted 3 to6-membered heterocycloalkyl), optionally substituted 6 to 10-memberedaryl, or optionally substituted 5 to 10-membered heteroaryl; and

n is an integer from 1 to 4.

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula IIa-2:

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula IIa-3:

wherein R⁴ and R⁵ are each independently selected from halogen, cyano,hydroxy, optionally substituted amine, optionally substituted amido,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 11-membered heterocycloalkyl (e.g., optionallysubstituted 3 to 6-membered heterocycloalkyl), optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl.

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula IIa-4:

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula IIa-5:

wherein X³ is N or CH;

m is 1 or 2;

R⁶, R⁷, R⁸, and R¹¹ are each independently selected from hydrogen,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 6-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl; or

R⁶ and R⁷ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered cycloalkyl or an optionallysubstituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R⁸ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R¹¹ combine with the atoms to which they are attached to form anoptionally substituted 4 to 8-membered heterocycloalkyl. In someembodiments, X³ is N. In some embodiments, m is 1. In some embodiments,R¹¹ is H. In some embodiments, X³ is N, m is 1, and R¹¹ is H.

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula IIa-6:

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula IIa-7:

In some embodiments (e.g., of any one of Formulae IIa-6 or IIa-7), R⁶ ismethyl.

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula IIa-8 or Formula IIa-9:

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula IIIa:

wherein a is 0 or 1.

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula IIIa-1:

wherein X² is N or CH;

each R³ is independently selected from halogen, cyano, hydroxy,optionally substituted amine, optionally substituted amido, optionallysubstituted C₁-C₆ alkyl, optionally substituted C₁-C₆ heteroalkyl,optionally substituted 3 to 6-membered cycloalkyl, optionallysubstituted 3 to 6-membered cycloalkenyl, optionally substituted 3 to11-membered heterocycloalkyl (e.g., optionally substituted 3 to6-membered heterocycloalkyl), optionally substituted 6 to 10-memberedaryl, or optionally substituted 5 to 10-membered heteroaryl; and

n is an integer from 1 to 4.

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula IIIa-2:

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula IIIa-3:

wherein R⁴ and R⁵ are each independently selected from halogen, cyano,hydroxy, optionally substituted amine, optionally substituted amido,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 11-membered heterocycloalkyl (e.g., optionallysubstituted 3 to 6-membered heterocycloalkyl), optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl.

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula IIIa-4:

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula IIIa-5:

wherein X³ is N or CH;

m is 1 or 2;

R⁶, R⁷, R⁸, and R¹¹ are each independently selected from hydrogen,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 6-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl; or

R⁶ and R⁷ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered cycloalkyl or an optionallysubstituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R⁸ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R¹¹ combine with the atoms to which they are attached to form anoptionally substituted 4 to 8-membered heterocycloalkyl. In someembodiments, X³ is N. In some embodiments, m is 1. In some embodiments,R¹¹ is hydrogen. In some embodiments, X³ is N, m is 1, and R¹¹ is H.

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula IIIa-6:

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula IIIa-7:

In some embodiments (e.g., of any one of Formulae IIIa-6 or IIIa-7), R⁶is methyl.

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula IIIa-8 or Formula IIIa-9:

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula IVa:

wherein R⁹ is H or C₁-C₆ alkyl; and

a is 0 or 1.

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula IVa-1:

wherein X² is N or CH;

each R³ is independently selected from halogen, cyano, hydroxy,optionally substituted amine, optionally substituted amido, optionallysubstituted C₁-C₆ alkyl, optionally substituted C₁-C₆ heteroalkyl,optionally substituted 3 to 6-membered cycloalkyl, optionallysubstituted 3 to 6-membered cycloalkenyl, optionally substituted 3 to11-membered heterocycloalkyl (e.g., optionally substituted 3 to6-membered heterocycloalkyl), optionally substituted 6 to 10-memberedaryl, or optionally substituted 5 to 10-membered heteroaryl; and

n is an integer from 1 to 4.

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula IVa-2:

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula IVa-3:

wherein R⁴ and R⁵ are each independently selected from halogen, cyano,hydroxy, optionally substituted amine, optionally substituted amido,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 11-membered heterocycloalkyl (e.g., optionallysubstituted 3 to 6-membered heterocycloalkyl), optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl.

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula IVa-4:

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula IVa-5:

wherein X³ is N or CH;

m is 1 or 2;

R⁶, R⁷, R⁸, and R¹¹ are each independently selected from hydrogen,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 6-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl; or

R⁶ and R⁷ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered cycloalkyl or an optionallysubstituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R⁸ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R¹¹ combine with the atoms to which they are attached to form anoptionally substituted 4 to 8-membered heterocycloalkyl. In someembodiments, X³ is N. In some embodiments, m is 1. In some embodiments,R¹¹ is hydrogen. In some embodiments, X³ is N, m is 1, and R¹¹ is H.

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula IVa-6:

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula IVa-7:

In some embodiments (e.g., of any one of Formulae IVa-6 or IVa-7), R⁶ ismethyl.

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula IVa-8 or Formula IVa-9:

In some embodiments (e.g., of any one of Formulae IVa, IVa-1, IVa-2,IVa-3, IVa-4, IVa-5, IVa-6, IVa-7, IVa-8, or IVa-9), R⁹ is methyl.

In some embodiments, Y is —NHS(O)₂— or —NHS(O)₂NH—.

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula Va:

wherein a is 0 or 1.

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula Va-1:

wherein X² is N or CH;

each R³ is independently selected from halogen, cyano, hydroxy,optionally substituted amine, optionally substituted amido, optionallysubstituted C₁-C₆ alkyl, optionally substituted C₁-C₆ heteroalkyl,optionally substituted 3 to 6-membered cycloalkyl, optionallysubstituted 3 to 6-membered cycloalkenyl, optionally substituted 3 to11-membered heterocycloalkyl (e.g., optionally substituted 3 to6-membered heterocycloalkyl), optionally substituted 6 to 10-memberedaryl, or optionally substituted 5 to 10-membered heteroaryl; and

n is an integer from 1 to 4.

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula Va-2:

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula Va-3:

wherein R⁴ and R⁵ are each independently selected from halogen, cyano,hydroxy, optionally substituted amine, optionally substituted amido,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 11-membered heterocycloalkyl (e.g., optionallysubstituted 3 to 6-membered heterocycloalkyl), optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl.

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula Va-4:

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula Va-5:

wherein X³ is N or CH;

m is 1 or 2;

R⁶, R⁷, R⁸, and R¹¹ are each independently selected from hydrogen,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 6-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl; or

R⁶ and R⁷ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered cycloalkyl or an optionallysubstituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R⁸ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R¹¹ combine with the atoms to which they are attached to form anoptionally substituted 4 to 8-membered heterocycloalkyl. In someembodiments, X³ is N. In some embodiments, m is 1. In some embodiments,R¹¹ is hydrogen. In some embodiments, X³ is N, m is 1, and R¹¹ is H.

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula VIa:

wherein a is 0 or 1.

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula VIa-1:

wherein X² is N or CH;

each R³ is independently selected from halogen, cyano, hydroxy,optionally substituted amine, optionally substituted amido, optionallysubstituted C₁-C₆ alkyl, optionally substituted C₁-C₆ heteroalkyl,optionally substituted 3 to 6-membered cycloalkyl, optionallysubstituted 3 to 6-membered cycloalkenyl, optionally substituted 3 to11-membered heterocycloalkyl (e.g., optionally substituted 3 to6-membered heterocycloalkyl), optionally substituted 6 to 10-memberedaryl, or optionally substituted 5 to 10-membered heteroaryl; and

n is an integer from 1 to 4.

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula VIa-2:

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula VIa-3:

wherein R⁴ and R⁵ are each independently selected from halogen, cyano,hydroxy, optionally substituted amine, optionally substituted amido,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 11-membered heterocycloalkyl (e.g., optionallysubstituted 3 to 6-membered heterocycloalkyl), optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl.

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula VIa-4:

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula VIa-5:

wherein X³ is N or CH;

m is 1 or 2;

R⁶, R⁷, R⁸, and R¹¹ are each independently selected from hydrogen,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 6-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl; or

R⁶ and R⁷ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered cycloalkyl or an optionallysubstituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R⁸ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R¹¹ combine with the atoms to which they are attached to form anoptionally substituted 4 to 8-membered heterocycloalkyl. In someembodiments, X³ is N. In some embodiments, m is 1. In some embodiments,R¹¹ is hydrogen. In some embodiments, X³ is N, m is 1, and R¹¹ is H.

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula VIIa:

wherein R⁹ is H or C₁-C₆ alkyl; and

a is 0 or 1.

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula VIIa-1:

wherein X² is N or CH;

each R³ is independently selected from halogen, cyano, hydroxy,optionally substituted amine, optionally substituted amido, optionallysubstituted C₁-C₆ alkyl, optionally substituted C₁-C₆ heteroalkyl,optionally substituted 3 to 6-membered cycloalkyl, optionallysubstituted 3 to 6-membered cycloalkenyl, optionally substituted 3 to11-membered heterocycloalkyl (e.g., optionally substituted 3 to6-membered heterocycloalkyl), optionally substituted 6 to 10-memberedaryl, or optionally substituted 5 to 10-membered heteroaryl; and

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula VIIa-2:

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula VIIa-3:

wherein R⁴ and R⁵ are each independently selected from halogen, cyano,hydroxy, optionally substituted amine, optionally substituted amido,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 11-membered heterocycloalkyl (e.g., optionallysubstituted 3 to 6-membered heterocycloalkyl), optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl.

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula VIIa-4:

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula VIIa-5:

wherein X³ is N or CH;

m is 1 or 2;

R⁶, R⁷, R⁸, and R¹¹ are each independently selected from hydrogen,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 6-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl; or

R⁶ and R⁷ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered cycloalkyl or an optionallysubstituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R⁸ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R¹¹ combine with the atoms to which they are attached to form anoptionally substituted 4 to 8-membered heterocycloalkyl. In someembodiments, X³ is N. In some embodiments, m is 1. In some embodiments,R¹¹ is hydrogen. In some embodiments, X³ is N, m is 1, and R¹¹ is H.

In some embodiments (e.g., of any one of Formulae VIIa, VIIa-1, VIIa-2,VIIa-3, VIIa-4, or VIIa-5), R⁹ is methyl.

In some embodiments, Y is —NHS(O)— or —NHS(O)NH—.

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula VIIIa:

wherein a is 0 or 1.

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula VIIIa-1:

wherein X² is N or CH;

each R³ is independently selected from halogen, cyano, hydroxy,optionally substituted amine, optionally substituted amido, optionallysubstituted C₁-C₆ alkyl, optionally substituted C₁-C₆ heteroalkyl,optionally substituted 3 to 6-membered cycloalkyl, optionallysubstituted 3 to 6-membered cycloalkenyl, optionally substituted 3 to11-membered heterocycloalkyl (e.g., optionally substituted 3 to6-membered heterocycloalkyl), optionally substituted 6 to 10-memberedaryl, or optionally substituted 5 to 10-membered heteroaryl; and

n is an integer from 1 to 4.

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula VIIIa-2:

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula VIIIa-3:

wherein R⁴ and R⁵ are each independently selected from halogen, cyano,hydroxy, optionally substituted amine, optionally substituted amido,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 11-membered heterocycloalkyl (e.g., optionallysubstituted 3 to 6-membered heterocycloalkyl), optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl.

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula VIIIa-4:

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula VIIIa-5:

wherein X³ is N or CH;

m is 1 or 2;

R⁶, R⁷, R⁸, and R¹¹ are each independently selected from hydrogen,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 6-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl; or

R⁶ and R⁷ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered cycloalkyl or an optionallysubstituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R⁸ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R¹¹ combine with the atoms to which they are attached to form anoptionally substituted 4 to 8-membered heterocycloalkyl. In someembodiments, X³ is N. In some embodiments, m is 1. In some embodiments,R¹¹ is hydrogen. In some embodiments, X³ is N, m is 1, and R¹¹ is H.

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula IXa:

wherein a is 0 or 1.

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula IXa-1:

wherein X² is N or CH;

each R³ is independently selected from halogen, cyano, hydroxy,optionally substituted amine, optionally substituted amido, optionallysubstituted C₁-C₆ alkyl, optionally substituted C₁-C₆ heteroalkyl,optionally substituted 3 to 6-membered cycloalkyl, optionallysubstituted 3 to 6-membered cycloalkenyl, optionally substituted 3 to11-membered heterocycloalkyl (e.g., optionally substituted 3 to6-membered heterocycloalkyl), optionally substituted 6 to 10-memberedaryl, or optionally substituted 5 to 10-membered heteroaryl; and

n is an integer from 1 to 4.

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula IXa-2:

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula IXa-3:

wherein R⁴ and R⁵ are each independently selected from halogen, cyano,hydroxy, optionally substituted amine, optionally substituted amido,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 11-membered heterocycloalkyl (e.g., optionallysubstituted 3 to 6-membered heterocycloalkyl), optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl.

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula IXa-4:

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula IXa-5:

wherein X³ is N or CH;

m is 1 or 2;

R⁶, R⁷, R⁸, and R¹¹ are each independently selected from hydrogen,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 6-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl; or

R⁶ and R⁷ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered cycloalkyl or an optionallysubstituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R⁸ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R¹¹ combine with the atoms to which they are attached to form anoptionally substituted 4 to 8-membered heterocycloalkyl. In someembodiments, X³ is N. In some embodiments, m is 1. In some embodiments,R¹¹ is hydrogen. In some embodiments, X³ is N, m is 1, and R¹¹ is H.

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula Xa:

wherein R⁹ is H or C₁-C₆ alkyl; and

a is 0 or 1.

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula Xa-1:

wherein X² is N or CH;

each R³ is independently selected from halogen, cyano, hydroxy,optionally substituted amine, optionally substituted amido, optionallysubstituted C₁-C₆ alkyl, optionally substituted C₁-C₆ heteroalkyl,optionally substituted 3 to 6-membered cycloalkyl, optionallysubstituted 3 to 6-membered cycloalkenyl, optionally substituted 3 to11-membered heterocycloalkyl (e.g., optionally substituted 3 to6-membered heterocycloalkyl), optionally substituted 6 to 10-memberedaryl, or optionally substituted 5 to 10-membered heteroaryl; and

n is an integer from 1 to 4.

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula Xa-2:

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula Xa-3:

wherein R⁴ and R⁵ are each independently selected from halogen, cyano,hydroxy, optionally substituted amine, optionally substituted amido,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 11-membered heterocycloalkyl (e.g., optionallysubstituted 3 to 6-membered heterocycloalkyl), optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl.

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula Xa-4:

In some embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the structure of Formula Xa-5:

wherein X³ is N or CH;

m is 1 or 2;

R⁶, R⁷, R⁸, and R¹¹ are each independently selected from hydrogen,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 6-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl; or

R⁶ and R⁷ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered cycloalkyl or an optionallysubstituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R⁸ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R¹¹ combine with the atoms to which they are attached to form anoptionally substituted 4 to 8-membered heterocycloalkyl. In someembodiments, X³ is N. In some embodiments, m is 1. In some embodiments,R¹¹ is hydrogen. In some embodiments, X³ is N, m is 1, and R¹¹ is H.

In some embodiments (e.g., of any one of Formulae Xa, Xa-1, Xa-2, Xa-3,Xa-4, or Xa-5), R⁹ is methyl.

In some embodiments of any aspect described herein, a is 0. In someembodiments of any of the above, a is 0.

In some embodiments of any aspect described herein, R² is optionallysubstituted C₁-C₆ alkyl. In some embodiments, R² is selected from—CH₂CH₃ or—CH₂CF₃.

In some embodiments of any aspect described herein, W is C₁-C₄ alkyl. Insome embodiments, W is:

In some embodiments of any aspect described herein, W is optionallysubstituted cyclopropyl, optionally substituted cyclobutyl, optionallysubstituted cyclopentyl, or optionally substituted cyclohexyl,optionally substituted piperidine, optionally substituted piperazine,optionally substituted pyridine, or optionally substituted phenyl.

In some embodiments of any aspect described herein, W is optionallysubstituted 3 to 10-membered heterocycloalkyl, optionally substituted 3to 10-membered cycloalkyl, optionally substituted 6 to 10-membered aryl,or optionally substituted 5 to 10-membered heteroaryl.

In some embodiments of any aspect described herein, W is optionallysubstituted 3 to 10-membered heterocycloalkyl. In some embodiments, W isselected from the following, or a stereoisomer thereof:

In some embodiments, W is selected from the following, or a stereoisomerthereof:

In some embodiments of any aspect described herein, W is optionallysubstituted 3 to 10-membered cycloalkyl. In some embodiments, W isselected from the following, or a stereoisomer thereof:

In some embodiments, W is selected from the following, or a stereoisomerthereof:

In some embodiments of any aspect described herein, W is optionallysubstituted 5 to 10-membered heteroaryl. In some embodiments, W isselected from the following, or a stereoisomer thereof:

In some embodiments of any aspect described herein, W is optionallysubstituted 6 to 10-membered aryl. In some embodiments, W is optionallysubstituted phenyl.

In some embodiments of any aspect described herein, W is optionallysubstituted C₁-C₃ heteroalkyl. In some embodiments, W is selected fromthe following, or a stereoisomer thereof:

In some embodiments, a compound of the present invention is selectedfrom Table 1a, or a pharmaceutically acceptable salt or stereoisomerthereof. In some embodiments, a compound of the present invention isselected from Table 1a, or a pharmaceutically acceptable salt oratropisomer thereof.

TABLE 1a Certain Compounds of the Present Invention Ex. # Structure A1

A2

A3

A4

A5

A6

A7

A8

A9

A10

A11

A12

A13

A14

A15

A16

A17

A18

A19

A20

A21

A22

A23

A24

A25

A26

A27

A28

A29

A30

A31

A32

A33

A34

A35

A36

A37

A38

A39

A40

A41

A42

A43

A44

A45

A46

A47

A48

A49

A50

A51

A52

A53

A54

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A59

A60

A61

A62

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A105

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A107

A108

A109

A110

A111

A112

A113

A114

A115

A116

A117

A118

A119

A120

A121

A122

A123

A124

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A126

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A128

A129

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A133

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A135

A136

A137

A138

A139

A140

A141

A142

A143

A144

A145

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A148

A149

A150

A151

A152

A153

A154

A155

A156

A157

A158

A159

A160

A161

A162

A163

A164

A165

A166

A167

A168

A169

A170

A171

A172

A173

A174

A175

A176

A177

A178

A179

A180

A181

A182

A183

A184

A185

A186

A187

A188

A189

A190

A191

A192

A193

A194

A195

A196

A197

A198

A199

A200

A201

A202

A203

A204

A205

A206

A207

A208

A209

A210

A211

A212

A213

A214

A215

A216

A217

A218

A219

A220

A221

A222

A223

A224

A225

A226

A227

A228

A229

A230

A231

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A238

A239

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A286

A287

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A290

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A292

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A301

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A328

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A341

A342

A343

A344

A345

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A347

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A349

A350

A351

A352

A353

Note that some compounds are shown with bonds as flat or wedged. In someinstances, the relative stereochemistry of stereoisomers has beendetermined; in some instances, the absolute stereochemistry has beendetermined. All stereoisomers of the compounds of the foregoing tableare contemplated by the present invention. In particular embodiments, anatropisomer of a compound of the foregoing table is contemplated. Anycompound shown in brackets indicates that the compound is adiastereomer, and the absolute stereochemistry of such diasteromer maynot be known.

In some embodiments, a compound of the present invention is selectedfrom Table 1b, or a pharmaceutically acceptable salt or stereoisomerthereof. In some embodiments, a compound of the present invention isselected from Table 1b, or a pharmaceutically acceptable salt oratropisomer thereof.

TABLE 1b Certain Compounds of the Present Invention Ex. # Structure A354

A355

A356

A357

A358

A359

A360

A361

A362

A363

A364

A365

A366

A367

A368

A369

A370

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A398

A399

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A405

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A419

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A427

A428

A429

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A477

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A531

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A541

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A592

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Note that some compounds are shown with bonds as flat or wedged. In someinstances, the relative stereochemistry of stereoisomers has beendetermined; in some instances, the absolute stereochemisty has beendetermined. All stereoisomers of the compounds of the foregoing tableare contemplated by the present invention. In particular embodiments, anatropsiomer of a compound of the foregoing table is contemplated. Anycompound shown in brackets indicatess that the compound is adiastereomer, and the absolute stereochemistry of such diastereomer maynot be known.

In some embodiments, a compound of the present invention is a compoundselected from Table 2, or a pharmaceutically acceptable salt orstereoisomer thereof. In some embodiments, a compound of the presentinvention is a compound selected from Table 2, or a pharmaceuticallyacceptable salt or atropisomer thereof

In some embodiments, a compound of the present invention is not acompound selected from Table 2. In some embodiments, a compound of thepresent invention is not a compound selected from Table 2, or apharmaceutically acceptable salt or stereoisomer thereof. In someembodiments, a compound of the present invention is not a compoundselected from Table 2, or a pharmaceutically acceptable salt oratropisomer thereof.

TABLE 2 Certain Compounds Ex. # Structure B1

B2

B3

B4

B5

B6

B7

B8

B9

B10

B11

B12

B13

B14

B15

B16

B17

B18

B19

B20

B21

B22

B23

B24

B25

In some embodiments, a compound of the present invention is a compoundselected from Table 3 (e.g., C1-C20 or C1-C21), or a pharmaceuticallyacceptable salt or stereoisomer thereof. In some embodiments, a compoundof the present invention is a compound selected from Table 3 (e.g.,C1-C20 or C1-C21), or a pharmaceutically acceptable salt or atropisomerthereof.

In some embodiments, a compound of the present invention is not acompound selected from Table 3 (e.g., C1-C20 or C1-C21). In someembodiments, a compound of the present invention is not a compoundselected from Table 3 (e.g., C1-C20 or C1-C21), or a pharmaceuticallyacceptable salt or stereoisomer thereof. In some embodiments, a compoundof the present invention is not a compound selected from Table 3 (e.g.,C1-C20 or C1-C21), or a pharmaceutically acceptable salt or atropisomerthereof.

TABLE 3 Certain Compounds Ex. # Structure C1

C2

C3

C4

C5

C6

C7

C8

C9

C10

C11

C12

C13

C14

C15

C16

C17

C18

C19

C20

C21

In some embodiments, a compound of the present invention has improvedoral bioavailability (% F) compared to what is known in the art. Methodsof measuring oral bioavailability are known in the art, and one suchmethod is provided below:

-   -   Oral bioavailability may be determined in BALB/c mice. Following        intravenous (IV) bolus and oral gavage (PO) administration of a        test compound, about 30 μL of whole blood samples are collected        at designated time points into tubes containing K₂EDTA. The        blood samples are centrifuged at 4600 rpm at 4° C. for about 5        minutes and plasma samples are stored at −80° C. prior to        bioanalysis. Plasma samples are extracted by protein        precipitation and analyzed by tandem mass spectrometry (LC        MS/MS) on, for example, an API 5500 system using electrospray        positive ionization.    -   All PK parameters may be derived from plasma concentration over        time data with noncompartment analysis using WinNonlin. The        bioavailability (F %, also % F) is estimated using the following        equation:

${F\mspace{14mu}\%} = {\frac{AUC_{\inf,{PO}}}{AUC_{\inf,{IV}}} \cdot \frac{Dose_{IV}}{Dose_{PO}}}$

-   -   AUC_(inf,PO) is the area under the plasma concentration overtime        from time zero to infinity following PO administration.    -   AUC_(inf,IV) is the area under the plasma concentration overtime        from time zero to infinity following IV administration.    -   Dose_(IV) is the total dose of IV administration    -   Dose_(PO) is the total dose of PO administration

In general, F % (or % F) values of over 30% are preferred, with valuesover 50% being more preferred.

In some embodiments, a compound of the present invention is selectivefor one or more particular Ras mutants over other Ras mutants orwild-type compared to what is known in the art. Methods of measuringsuch selectivity are known in the art, such as the Ras-Raf bindingassay, a protocol for which is provided in the Examples below.Accordingly, in some embodiments, compounds of the present invention areselective for KRAS^(G12C) over other Ras mutants or over wild-type. Insome embodiments, compounds of the present invention are selective forKRAS^(G12D) over other Ras mutants or over wild-type. In someembodiments, compounds of the present invention are selective forKRAS^(G12V) over other Ras mutants or over wild-type. In someembodiments, compounds of the present invention are selective forKRAS^(G12D) over other Ras mutants or over wild-type. In someembodiments, compounds of the present invention are selective forNRAS^(Q61K) over other Ras mutants or over wild-type. In someembodiments, compounds of the present invention are selective forKRAS^(G12D) and KRAS^(G12V) over other Ras mutants and wild-type.Compounds of the present invention may also exhibit greater selectivitywith respect to other RAS mutants disclosed herein, or combinationsthereof. In some embodiments, compounds of the present invention exhibitan 1C₅₀ value of less than 30 nm for one or more Ras mutants describedherein in the Ras-Raf binding assay described above.

In some embodiments, a compound of the present invention is more potentfor one or more particular Ras mutants over other Ras mutants orwild-type compared to what is known in the art. Methods of measuringsuch potency are known in the art, such as the pERK assay, a protocolfor which is provided in the Examples below. Accordingly, in someembodiments, compounds of the present invention exhibit greater potencywith respect to KRAS^(G12D) than what is known in the art. In someembodiments, compounds of the present invention exhibit greater potencywith respect to KRAS^(G12V) than what is known in the art. In someembodiments, compounds of the present invention exhibit greater potencywith respect to KRAS^(G12C) than what is known in the art. In someembodiments, compounds of the present invention exhibit greater potencywith respect to both KRAS^(G12D) and KRAS^(G12V) than what is known inthe art. Compounds of the present invention may also exhibit greaterpotency with respect to other RAS mutants disclosed herein, orcombinations thereof.

In some embodiments, a compound of the present invention exhibits agreater detrimental effect on cell viability with respect to one or moreparticular Ras mutants over other Ras mutants or wild-type compared towhat is known in the art. Methods of measuring cell viability are knownin the art, such as the CellTiter-Glo® Cell Viability Assay assay, aprotocol for which is provided in the Examples below. Accordingly, insome embodiments, compounds of the present invention exhibit a greaterdecrease in cell viability with respect to KRAS^(G12D) compared to whatis known in the art. In some embodiments, compounds of the presentinvention exhibit a greater decrease in cell viability with respect toKRAS^(G12V) compared to what is known in the art. In some embodiments,compounds of the present invention exhibit a greater decrease in cellviability with respect to KRAS^(G12C) compared to what is known in theart. In some embodiments, compounds of the present invention exhibit agreater decrease in cell viability with respect to both KRAS^(G12D) andKRAS^(G12V) compared to what is known in the art. Compounds of thepresent invention may also exhibit a greater decrease in cell viabilityrespect to other RAS mutants disclosed herein, or combinations thereof.

In some embodiments, a compound of the present invention may exhibitgreater metabolic stability, permeability, or solubility, or acombination thereof, versus what is known in the art. Methods formeasuring such properties are known in the art. In some embodiments, acompound of the present invention may exhibit improvements with respectto any of the following properties, or a combination thereof, comparedto what is known in the art: selectivity, potency, cell viability,metabolic stability, permeability, or solubility.

In some embodiments, a compound of the present invention is or acts as aprodrug, such as with respect to administration to a cell or to asubject in need thereof.

Also provided are pharmaceutical compositions comprising a compound ofthe present invention, or a pharmaceutically acceptable salt thereof,and a pharmaceutically acceptable excipient.

Further provided is a method of treating cancer in a subject in needthereof, the method comprising administering to the subject atherapeutically effective amount of a compound of the present invention,or a pharmaceutically acceptable salt thereof. The cancer may, forexample, be pancreatic cancer, colorectal cancer, non-small cell lungcancer, acute myeloid leukemia, multiple myeloma, thyroid glandadenocarcinoma, a myelodysplastic syndrome, or squamous cell lungcarcinoma. In some embodiments, the cancer comprises a Ras mutation,such as K-Ras G12C, K-Ras G12D, K-Ras G12V, K-Ras G12S, K-Ras G13C,K-Ras G13D, K-Ras Q61H, K-Ras Q61R, K-Ras Q61K, or K-Ras Q61L, or acombination thereof. In some embodiments, the cancer comprises a Rasmutation, such as N-Ras G12D, N-Ras Q61R, N-Ras Q61K, N-Ras Q61L, N-RasQ61H, or N-Ras Q61P, or a combination thereof. Other Ras mutations aredescribed herein.

Further provided is a method of treating a Ras protein-related disorderin a subject in need thereof, the method comprising administering to thesubject a therapeutically effective amount of a compound of the presentinvention, or a pharmaceutically acceptable salt thereof.

Further provided is a method of inhibiting a Ras protein in a cell, themethod comprising contacting the cell with an effective amount of acompound of the present invention, or a pharmaceutically acceptable saltthereof. For example, the Ras protein is K-Ras G12C, K-Ras G12D, K-RasG12V, K-Ras G12S, K-Ras G13C, K-Ras G13D, K-Ras Q61H, K-Ras Q61R, K-RasQ61K, or K-Ras Q61L. The Ras protein may be, for example, N-Ras G12D,N-Ras Q61R, N-Ras Q61K, N-Ras Q61L, N-Ras Q61H, or N-Ras Q61P. Other Rasproteins are described herein. The cell may be a cancer cell, such as apancreatic cancer cell, a colorectal cancer cell, a lung cancer (e.g.,non-small cell lung cancer cell), an acute myeloid leukemia cell, amultiple myeloma cell, a thyroid gland adenocarcinoma cell, amyelodysplastic syndrome cell, a melanoma cell, or a squamous cell lungcarcinoma cell. Other cancer types are described herein. The cell may bein vivo or in vitro.

With respect to compounds of the present invention, one stereoisomer mayexhibit better inhibition than another stereoisomer. For example, oneatropisomer may exhibit inhibition, whereas the other atropisomer mayexhibit little or no inhibition.

In some embodiments, a method or use described herein further comprisesadministering an additional anti-cancer therapy. In some embodiments,the additional anti-cancer therapy is a HER2 inhibitor, an EGFRinhibitor, a second Ras inhibitor, a SHP2 inhibitor, a SOS1 inhibitor, aRaf inhibitor, a MEK inhibitor, an ERK inhibitor, a PI3K inhibitor, aPTEN inhibitor, an AKT inhibitor, an mTORC1 inhibitor, a BRAF inhibitor,a PD-L1 inhibitor, a PD-1 inhibitor, a CDK4/6 inhibitor, or acombination thereof. In some embodiments, the additional anticancertherapy is a SHP2 inhibitor. Other additional anti-cancer therapies aredescribed herein.

Methods of Synthesis

The compounds described herein may be made from commercially availablestarting materials or synthesized using known organic, inorganic, orenzymatic processes.

The compounds of the present invention can be prepared in a number ofways well known to those skilled in the art of organic synthesis. By wayof example, compounds of the present invention can be synthesized usingthe methods described in the Schemes below, together with syntheticmethods known in the art of synthetic organic chemistry, or variationsthereon as appreciated by those skilled in the art. These methodsinclude but are not limited to those methods described in the Schemesbelow.

A general synthesis of macrocyclic esters is outlined in Scheme 1. Anappropriately substituted indolyl boronic ester (1) can be prepared infour steps starting from protected3-(5-bromo-2-iodo-1H-indol-3-yl)-2,2-dimethylpropan-1-ol andappropriately substituted boronic acid, including palladium mediatedcoupling, alkylation, de-protection, and palladium mediated borylationreactions.

Methyl-amino-3-(4-bromothiazol-2-yl)propanoyl)hexahydropyridazine-3-carboxylate(3) can be prepared via coupling of(S)-2-amino-3-(4-bromothiazol-2-yl)propanoic acid (2) with methyl(S)-hexahydropyridazine-3-carboxylate.

The final macrocyclic esters can be made by coupling ofmethyl-amino-3-(4-bromothiazol-2-yl)propanoyl)hexahydropyridazine-3-carboxylate(3) and an appropriately substituted indolyl boronic ester (1) in thepresence of Pd catalyst followed by hydrolysis and macrolactonizationsteps to result in an appropriately protected macrocyclic intermediate(5). Deprotection and coupling with an appropriately substitutedcarboxylic acid (or other coupling partner) can result in a macrocyclicproduct. Additional deprotection or functionalization steps could berequired to produce a final compound 6.

Further, with respect to Scheme 1, the thiazole may be replaced with analternative optionally substituted 5 to 6-membered heteroarylene, or anoptionally substituted 3 to 6-membered cycloalkylene, optionallysubstituted 3 to 6-membered heterocycloalkylene (e.g., morpholino), oroptionally substituted 6-membered arylene (e.g., phenyl).

Alternatively, macrocyclic esters can be prepared as described in Scheme2. An appropriately substituted and protected indolyl boronic ester (7)can be coupled in the presence of Pd catalyst with(S)-2-amino-3-(4-bromothiazol-2-yl)propanoic acid, followed byiodination, deprotection, and ester hydrolysis. Subsequent coupling withmethyl (S)-hexahydropyridazine-3-carboxylate, followed by hydrolysis andmacrolactonization can result in iodo intermediate (11). Subsequentpalladium mediated borylation and coupling in the presence of Pdcatalyst with an appropriately substituted iodo aryl or iodo heteroarylintermediate can yield an appropriately protected macrocyclicintermediate. Alkylation, deprotection and coupling with anappropriately substituted carboxylic acid carboxylic acid (or othercoupling partner) results in a macrocyclic product. Additionaldeprotection or functionalization steps could be required to produce afinal compound 6.

Further, with respect to Scheme 2, the thiazole may be replaced with analternative optionally substituted 5 to 6-membered heteroarylene, or anoptionally substituted 3 to 6-membered cycloalkylene, optionallysubstituted 3 to 6-membered heterocycloalkylene (e.g., morpholino), oroptionally substituted 6-membered arylene (e.g., phenyl).

Compounds of Table 1a or Table 1b herein were prepared using methodsdisclosed herein or were prepared using methods described hereincombined with the knowledge of one of skill in the art.

Pharmaceutical Compositions and Methods of Use PharmaceuticalCompositions and Methods of Administration

The compounds with which the invention is concerned are Ras inhibitors,and are useful in the treatment of cancer. Accordingly, one embodimentof the present invention provides pharmaceutical compositions containinga compound of the invention or a pharmaceutically acceptable saltthereof, and a pharmaceutically acceptable excipient, as well as methodsof using the compounds of the invention to prepare such compositions.

As used herein, the term “pharmaceutical composition” refers to acompound, such as a compound of the present invention, or apharmaceutically acceptable salt thereof, formulated together with apharmaceutically acceptable excipient.

In some embodiments, a compound is present in a pharmaceuticalcomposition in unit dose amount appropriate for administration in atherapeutic regimen that shows a statistically significant probabilityof achieving a predetermined therapeutic effect when administered to arelevant population. In some embodiments, pharmaceutical compositionsmay be specially formulated for administration in solid or liquid form,including those adapted for the following: oral administration, forexample, drenches (aqueous or non-aqueous solutions or suspensions),tablets, e.g., those targeted for buccal, sublingual, and systemicabsorption, boluses, powders, granules, pastes for application to thetongue; parenteral administration, for example, by subcutaneous,intramuscular, intravenous or epidural injection as, for example, asterile solution or suspension, or sustained-release formulation;topical application, for example, as a cream, ointment, or acontrolled-release patch or spray applied to the skin, lungs, or oralcavity; intravaginally or intrarectally, for example, as a pessary,cream, or foam; sublingually; ocularly; transdermally; or nasally,pulmonary, and to other mucosal surfaces.

A “pharmaceutically acceptable excipient,” as used herein, refers anyinactive ingredient (for example, a vehicle capable of suspending ordissolving the active compound) having the properties of being nontoxicand non-inflammatory in a subject. Typical excipients include, forexample: antiadherents, antioxidants, binders, coatings, compressionaids, d is integrants, dyes (colors), emollients, emulsifiers, fillers(diluents), film formers or coatings, flavors, fragrances, glidants(flow enhancers), lubricants, preservatives, printing inks, sorbents,suspensing or dispersing agents, sweeteners, or waters of hydration.Excipients include, but are not limited to: butylated optionallysubstituted hydroxyltoluene (BHT), calcium carbonate, calcium phosphate(dibasic), calcium stearate, croscarmellose, crosslinked polyvinylpyrrolidone, citric acid, crospovidone, cysteine, ethylcellulose,gelatin, optionally substituted hydroxylpropyl cellulose, optionallysubstituted hydroxyl propyl methylcellulose, lactose, magnesiumstearate, maltitol, mannitol, methionine, methylcellulose, methylparaben, microcrystalline cellulose, polyethylene glycol, polyvinylpyrrolidone, povidone, pregelatinized starch, propyl paraben, retinylpalmitate, shellac, silicon dioxide, sodium carboxymethyl cellulose,sodium citrate, sodium starch glycolate, sorbitol, starch (corn),stearic acid, stearic acid, sucrose, talc, titanium dioxide, vitamin A,vitamin E, vitamin C, and xylitol. Those of ordinary skill in the artare familiar with a variety of agents and materials useful asexcipients. See, e.g., e.g., Ansel, et al., Ansel's PharmaceuticalDosage Forms and Drug Delivery Systems. Philadelphia: Lippincott,Williams & Wilkins, 2004; Gennaro, et al., Remington: The Science andPractice of Pharmacy. Philadelphia: Lippincott, Williams & Wilkins,2000; and Rowe, Handbook of Pharmaceutical Excipients. Chicago,Pharmaceutical Press, 2005. In some embodiments, a composition includesat least two different pharmaceutically acceptable excipients.

Compounds described herein, whether expressly stated or not, may beprovided or utilized in salt form, e.g., a pharmaceutically acceptablesalt form, unless expressly stated to the contrary. The term“pharmaceutically acceptable salt,” as use herein, refers to those saltsof the compounds described herein that are, within the scope of soundmedical judgment, suitable for use in contact with the tissues of humansand other animals without undue toxicity, irritation, allergic responseand the like, and are commensurate with a reasonable benefit/risk ratio.Pharmaceutically acceptable salts are well known in the art. Forexample, pharmaceutically acceptable salts are described in: Berge etal., J. Pharmaceutical Sciences 66:1-19, 1977 and in PharmaceuticalSalts: Properties, Selection, and Use, (Eds. P. H. Stahl and C. G.Wermuth), Wiley-VCH, 2008. The salts can be prepared in situ during thefinal isolation and purification of the compounds described herein orseparately by reacting the free base group with a suitable organic acid.

The compounds of the invention may have ionizable groups so as to becapable of preparation as pharmaceutically acceptable salts. These saltsmay be acid addition salts involving inorganic or organic acids or thesalts may, in the case of acidic forms of the compounds of theinvention, be prepared from inorganic or organic bases. In someembodiments, the compounds are prepared or used as pharmaceuticallyacceptable salts prepared as addition products of pharmaceuticallyacceptable acids or bases. Suitable pharmaceutically acceptable acidsand bases are well-known in the art, such as hydrochloric, sulfuric,hydrobromic, acetic, lactic, citric, or tartaric acids for forming acidaddition salts, and potassium hydroxide, sodium hydroxide, ammoniumhydroxide, caffeine, various amines, and the like for forming basicsalts. Methods for preparation of the appropriate salts arewell-established in the art.

Representative acid addition salts include acetate, adipate, alginate,ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate,butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate,digluconate, dodecylsulfate, ethanesulfonate, fumarate, glucoheptonate,glycerophosphate, hemisulfate, heptonate, hexanoate, hydrobromide,hydrochloride, hydroiodide, 2-optionally substitutedhydroxyl-ethanesulfonate, lactobionate, lactate, laurate, laurylsulfate, malate, maleate, malonate, methanesulfonate,2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate,pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate,pivalate, propionate, stearate, succinate, sulfate, tartrate,thiocyanate, toluenesulfonate, undecanoate, valerate salts and the like.Representative alkali or alkaline earth metal salts include sodium,lithium, potassium, calcium, magnesium and the like, as well as nontoxicammonium, quaternary ammonium, and amine cations, including, but notlimited to ammonium, tetramethylammonium, tetraethylammonium,methylamine, dimethylamine, trimethylamine, triethylamine, ethylamineand the like.

As used herein, the term “subject” refers to any member of the animalkingdom. In some embodiments, “subject” refers to humans, at any stageof development. In some embodiments, “subject” refers to a humanpatient. In some embodiments, “subject” refers to non-human animals. Insome embodiments, the non-human animal is a mammal (e.g., a rodent, amouse, a rat, a rabbit, a monkey, a dog, a cat, a sheep, cattle, aprimate, or a pig). In some embodiments, subjects include, but are notlimited to, mammals, birds, reptiles, amphibians, fish, or worms. Insome embodiments, a subject may be a transgenic animal,genetically-engineered animal, or a clone.

As used herein, the term “dosage form” refers to a physically discreteunit of a compound (e.g., a compound of the present invention) foradministration to a subject. Each unit contains a predetermined quantityof compound. In some embodiments, such quantity is a unit dosage amount(or a whole fraction thereof) appropriate for administration inaccordance with a dosing regimen that has been determined to correlatewith a desired or beneficial outcome when administered to a relevantpopulation (i.e., with a therapeutic dosing regimen). Those of ordinaryskill in the art appreciate that the total amount of a therapeuticcomposition or compound administered to a particular subject isdetermined by one or more attending physicians and may involveadministration of multiple dosage forms. As used herein, the term“dosing regimen” refers to a set of unit doses (typically more than one)that are administered individually to a subject, typically separated byperiods of time. In some embodiments, a given therapeutic compound(e.g., a compound of the present invention) has a recommended dosingregimen, which may involve one or more doses. In some embodiments, adosing regimen comprises a plurality of doses each of which areseparated from one another by a time period of the same length; in someembodiments, a dosing regimen comprises a plurality of doses and atleast two different time periods separating individual doses. In someembodiments, all doses within a dosing regimen are of the same unit doseamount. In some embodiments, different doses within a dosing regimen areof different amounts. In some embodiments, a dosing regimen comprises afirst dose in a first dose amount, followed by one or more additionaldoses in a second dose amount different from the first dose amount. Insome embodiments, a dosing regimen comprises a first dose in a firstdose amount, followed by one or more additional doses in a second doseamount same as the first dose amount. In some embodiments, a dosingregimen is correlated with a desired or beneficial outcome whenadministered across a relevant population (i.e., is a therapeutic dosingregimen).

A “therapeutic regimen” refers to a dosing regimen whose administrationacross a relevant population is correlated with a desired or beneficialtherapeutic outcome.

The term “treatment” (also “treat” or “treating”), in its broadestsense, refers to any administration of a substance (e.g., a compound ofthe present invention) that partially or completely alleviates,ameliorates, relieves, inhibits, delays onset of, reduces severity of,or reduces incidence of one or more symptoms, features, or causes of aparticular disease, disorder, or condition. In some embodiments, suchtreatment may be administered to a subject who does not exhibit signs ofthe relevant disease, disorder or condition or of a subject who exhibitsonly early signs of the disease, disorder, or condition. Alternatively,or additionally, in some embodiments, treatment may be administered to asubject who exhibits one or more established signs of the relevantdisease, disorder or condition. In some embodiments, treatment may be ofa subject who has been diagnosed as suffering from the relevant disease,disorder, or condition. In some embodiments, treatment may be of asubject known to have one or more susceptibility factors that arestatistically correlated with increased risk of development of therelevant disease, disorder, or condition.

The term “therapeutically effective amount” means an amount that issufficient, when administered to a population suffering from orsusceptible to a disease, disorder, or condition in accordance with atherapeutic dosing regimen, to treat the disease, disorder, orcondition. In some embodiments, a therapeutically effective amount isone that reduces the incidence or severity of, or delays onset of, oneor more symptoms of the disease, disorder, or condition. Those ofordinary skill in the art will appreciate that the term “therapeuticallyeffective amount” does not in fact require successful treatment beachieved in a particular individual. Rather, a therapeutically effectiveamount may be that amount that provides a particular desiredpharmacological response in a significant number of subjects whenadministered to patients in need of such treatment. It is specificallyunderstood that particular subjects may, in fact, be “refractory” to a“therapeutically effective amount.” In some embodiments, reference to atherapeutically effective amount may be a reference to an amount asmeasured in one or more specific tissues (e.g., a tissue affected by thedisease, disorder or condition) or fluids (e.g., blood, saliva, serum,sweat, tears, urine). Those of ordinary skill in the art will appreciatethat, in some embodiments, a therapeutically effective amount may beformulated or administered in a single dose. In some embodiments, atherapeutically effective amount may be formulated or administered in aplurality of doses, for example, as part of a dosing regimen.

For use as treatment of subjects, the compounds of the invention, or apharmaceutically acceptable salt thereof, can be formulated aspharmaceutical or veterinary compositions. Depending on the subject tobe treated, the mode of administration, and the type of treatmentdesired, e.g., prevention, prophylaxis, or therapy, the compounds, or apharmaceutically acceptable salt thereof, are formulated in waysconsonant with these parameters. A summary of such techniques may befound in Remington: The Science and Practice of Pharmacy, 21^(st)Edition, Lippincott Williams & Wilkins, (2005); and Encyclopedia ofPharmaceutical Technology, eds. J. Swarbrick and J. C. Boylan,1988-1999, Marcel Dekker, New York, each of which is incorporated hereinby reference.

Compositions can be prepared according to conventional mixing,granulating or coating methods, respectively, and the presentpharmaceutical compositions can contain from about 0.1% to about 99%,from about 5% to about 90%, or from about 1% to about 20% of a compoundof the present invention, or pharmaceutically acceptable salt thereof,by weight or volume. In some embodiments, compounds, or apharmaceutically acceptable salt thereof, described herein may bepresent in amounts totaling 1-95% by weight of the total weight of acomposition, such as a pharmaceutical composition.

The composition may be provided in a dosage form that is suitable forintraarticular, oral, parenteral (e.g., intravenous, intramuscular),rectal, cutaneous, subcutaneous, topical, transdermal, sublingual,nasal, vaginal, intravesicular, intraurethral, intrathecal, epidural,aural, or ocular administration, or by injection, inhalation, or directcontact with the nasal, genitourinary, reproductive or oral mucosa.Thus, the pharmaceutical composition may be in the form of, e.g.,tablets, capsules, pills, powders, granulates, suspensions, emulsions,solutions, gels including hydrogels, pastes, ointments, creams,plasters, drenches, osmotic delivery devices, suppositories, enemas,injectables, implants, sprays, preparations suitable for iontophoreticdelivery, or aerosols. The compositions may be formulated according toconventional pharmaceutical practice.

As used herein, the term “administration” refers to the administrationof a composition (e.g., a compound, or a preparation that includes acompound as described herein) to a subject or system. Administration toan animal subject (e.g., to a human) may be by any appropriate route.For example, in some embodiments, administration may be bronchial(including by bronchial instillation), buccal, enteral, interdermal,intra-arterial, intradermal, intragastric, intramedullary,intramuscular, intranasal, intraperitoneal, intrathecal, intravenous,intraventricular, mucosal, nasal, oral, rectal, subcutaneous,sublingual, topical, tracheal (including by intratracheal instillation),transdermal, vaginal or vitreal.

Formulations may be prepared in a manner suitable for systemicadministration or topical or local administration. Systemic formulationsinclude those designed for injection (e.g., intramuscular, intravenousor subcutaneous injection) or may be prepared for transdermal,transmucosal, or oral administration. A formulation will generallyinclude a diluent as well as, in some cases, adjuvants, buffers,preservatives and the like. Compounds, or a pharmaceutically acceptablesalt thereof, can be administered also in liposomal compositions or asmicroemulsions.

For injection, formulations can be prepared in conventional forms asliquid solutions or suspensions or as solid forms suitable for solutionor suspension in liquid prior to injection or as emulsions. Suitableexcipients include, for example, water, saline, dextrose, glycerol andthe like. Such compositions may also contain amounts of nontoxicauxiliary substances such as wetting or emulsifying agents, pH bufferingagents and the like, such as, for example, sodium acetate, sorbitanmonolaurate, and so forth.

Various sustained release systems for drugs have also been devised. See,for example, U.S. Pat. No. 5,624,677.

Systemic administration may also include relatively noninvasive methodssuch as the use of suppositories, transdermal patches, transmucosaldelivery and intranasal administration. Oral administration is alsosuitable for compounds of the invention, or a pharmaceuticallyacceptable salt thereof. Suitable forms include syrups, capsules, andtablets, as is understood in the art.

Each compound, or a pharmaceutically acceptable salt thereof, asdescribed herein, may be formulated in a variety of ways that are knownin the art. For example, the first and second agents of the combinationtherapy may be formulated together or separately. Other modalities ofcombination therapy are described herein.

The individually or separately formulated agents can be packagedtogether as a kit. Non-limiting examples include, but are not limitedto, kits that contain, e.g., two pills, a pill and a powder, asuppository and a liquid in a vial, two topical creams, etc. The kit caninclude optional components that aid in the administration of the unitdose to subjects, such as vials for reconstituting powder forms,syringes for injection, customized IV delivery systems, inhalers, etc.Additionally, the unit dose kit can contain instructions for preparationand administration of the compositions. The kit may be manufactured as asingle use unit dose for one subject, multiple uses for a particularsubject (at a constant dose or in which the individual compounds, or apharmaceutically acceptable salt thereof, may vary in potency as therapyprogresses); or the kit may contain multiple doses suitable foradministration to multiple subjects (“bulk packaging”). The kitcomponents may be assembled in cartons, blister packs, bottles, tubes,and the like.

Formulations for oral use include tablets containing the activeingredient(s) in a mixture with non-toxic pharmaceutically acceptableexcipients. These excipients may be, for example, inert diluents orfillers (e.g., sucrose, sorbitol, sugar, mannitol, microcrystallinecellulose, starches including potato starch, calcium carbonate, sodiumchloride, lactose, calcium phosphate, calcium sulfate, or sodiumphosphate); granulating and disintegrating agents (e.g., cellulosederivatives including microcrystalline cellulose, starches includingpotato starch, croscarmellose sodium, alginates, or alginic acid);binding agents (e.g., sucrose, glucose, sorbitol, acacia, alginic acid,sodium alginate, gelatin, starch, pregelatinized starch,microcrystalline cellulose, magnesium aluminum silicate,carboxymethylcellulose sodium, methylcellulose, optionally substitutedhydroxylpropyl methylcellulose, ethylcellulose, polyvinylpyrrolidone, orpolyethylene glycol); and lubricating agents, glidants, andantiadhesives (e.g., magnesium stearate, zinc stearate, stearic acid,silicas, hydrogenated vegetable oils, or talc). Other pharmaceuticallyacceptable excipients can be colorants, flavoring agents, plasticizers,humectants, buffering agents, and the like.

Two or more compounds may be mixed together in a tablet, capsule, orother vehicle, or may be partitioned. In one example, the first compoundis contained on the inside of the tablet, and the second compound is onthe outside, such that a substantial portion of the second compound isreleased prior to the release of the first compound.

Formulations for oral use may also be provided as chewable tablets, oras hard gelatin capsules wherein the active ingredient is mixed with aninert solid diluent (e.g., potato starch, lactose, microcrystallinecellulose, calcium carbonate, calcium phosphate or kaolin), or as softgelatin capsules wherein the active ingredient is mixed with water or anoil medium, for example, peanut oil, liquid paraffin, or olive oil.Powders, granulates, and pellets may be prepared using the ingredientsmentioned above under tablets and capsules in a conventional mannerusing, e.g., a mixer, a fluid bed apparatus or a spray drying equipment.

Dissolution or diffusion-controlled release can be achieved byappropriate coating of a tablet, capsule, pellet, or granulateformulation of compounds, or by incorporating the compound, or apharmaceutically acceptable salt thereof, into an appropriate matrix. Acontrolled release coating may include one or more of the coatingsubstances mentioned above or, e.g., shellac, beeswax, glycowax, castorwax, carnauba wax, stearyl alcohol, glyceryl monostearate, glyceryldistearate, glycerol palmitostearate, ethylcellulose, acrylic resins,dl-polylactic acid, cellulose acetate butyrate, polyvinyl chloride,polyvinyl acetate, vinyl pyrrolidone, polyethylene, polymethacrylate,methylmethacrylate, 2-optionally substituted hydroxylmethacrylate,methacrylate hydrogels, 1,3 butylene glycol, ethylene glycolmethacrylate, or polyethylene glycols. In a controlled release matrixformulation, the matrix material may also include, e.g., hydratedmethylcellulose, carnauba wax and stearyl alcohol, carbopol 934,silicone, glyceryl tristearate, methyl acrylate-methyl methacrylate,polyvinyl chloride, polyethylene, or halogenated fluorocarbon.

The liquid forms in which the compounds, or a pharmaceuticallyacceptable salt thereof, and compositions of the present invention canbe incorporated for administration orally include aqueous solutions,suitably flavored syrups, aqueous or oil suspensions, and flavoredemulsions with edible oils such as cottonseed oil, sesame oil, coconutoil, or peanut oil, as well as elixirs and similar pharmaceuticalvehicles.

Generally, when administered to a human, the oral dosage of any of thecompounds of the invention, or a pharmaceutically acceptable saltthereof, will depend on the nature of the compound, and can readily bedetermined by one skilled in the art. A dosage may be, for example,about 0.001 mg to about 2000 mg per day, about 1 mg to about 1000 mg perday, about 5 mg to about 500 mg per day, about 100 mg to about 1500 mgper day, about 500 mg to about 1500 mg per day, about 500 mg to about2000 mg per day, or any range derivable therein. In some embodiments,the daily dose range for oral administration, for example, may liewithin the range of from about 0.001 mg to about 2000 mg per kg bodyweight of a human, in single or divided doses. On the other hand, it maybe necessary to use dosages outside these limits in some cases.

In some embodiments, the pharmaceutical composition may further comprisean additional compound having antiproliferative activity. Depending onthe mode of administration, compounds, or a pharmaceutically acceptablesalt thereof, will be formulated into suitable compositions to permitfacile delivery. Each compound, or a pharmaceutically acceptable saltthereof, of a combination therapy may be formulated in a variety of waysthat are known in the art. For example, the first and second agents ofthe combination therapy may be formulated together or separately.Desirably, the first and second agents are formulated together for thesimultaneous or near simultaneous administration of the agents.

It will be appreciated that the compounds and pharmaceuticalcompositions of the present invention can be formulated and employed incombination therapies, that is, the compounds and pharmaceuticalcompositions can be formulated with or administered concurrently with,prior to, or subsequent to, one or more other desired therapeutics ormedical procedures. The particular combination of therapies(therapeutics or procedures) to employ in a combination regimen willtake into account compatibility of the desired therapeutics orprocedures and the desired therapeutic effect to be achieved. It willalso be appreciated that the therapies employed may achieve a desiredeffect for the same disorder, or they may achieve different effects(e.g., control of any adverse effects).

Administration of each drug in a combination therapy, as describedherein, can, independently, be one to four times daily for one day toone year, and may even be for the life of the subject. Chronic,long-term administration may be indicated.

Methods of Use

In some embodiments, the invention discloses a method of treating adisease or disorder that is characterized by aberrant Ras activity dueto a Ras mutant. In some embodiments, the disease or disorder is acancer.

Accordingly, also provided is a method of treating cancer in a subjectin need thereof, the method comprising administering to the subject atherapeutically effective amount of a compound of the present invention,or a pharmaceutically acceptable salt thereof, or a pharmaceuticalcomposition comprising such a compound or salt. In some embodiments, thecancer is colorectal cancer, non-small cell lung cancer, small-cell lungcancer, pancreatic cancer, appendiceal cancer, melanoma, acute myeloidleukemia, small bowel cancer, ampullary cancer, germ cell cancer,cervical cancer, cancer of unknown primary origin, endometrial cancer,esophagogastric cancer, GI neuroendocrine cancer, ovarian cancer, sexcord stromal tumor cancer, hepatobiliary cancer, or bladder cancer. Insome embodiments, the cancer is appendiceal, endometrial or melanoma.Also provided is a method of treating a Ras protein-related disorder ina subject in need thereof, the method comprising administering to thesubject a therapeutically effective amount of a compound of the presentinvention, or a pharmaceutically acceptable salt thereof, or apharmaceutical composition comprising such a compound or salt.

In some embodiments, the compounds of the present invention orpharmaceutically acceptable salts thereof, pharmaceutical compositionscomprising such compounds or salts, and methods provided herein may beused for the treatment of a wide variety of cancers including tumorssuch as lung, prostate, breast, brain, skin, cervical carcinomas,testicular carcinomas, etc. More particularly, cancers that may betreated by the compounds or salts thereof, pharmaceutical compositionscomprising such compounds or salts, and methods of the inventioninclude, but are not limited to tumor types such as astrocytic, breast,cervical, colorectal, endometrial, esophageal, gastric, head and neck,hepatocellular, laryngeal, lung, oral, ovarian, prostate and thyroidcarcinomas and sarcomas. Other cancers include, for example:

-   -   Cardiac, for example: sarcoma (angiosarcoma, fibrosarcoma,        rhabdomyosarcoma, liposarcoma), myxoma, rhabdomyoma, fibroma,        lipoma and teratoma;    -   Lung, for example: bronchogenic carcinoma (squamous cell,        undifferentiated small cell, undifferentiated large cell,        adenocarcinoma), alveolar (bronchiolar) carcinoma, bronchial        adenoma, sarcoma, lymphoma, chondromatous hamartoma,        mesothelioma;    -   Gastrointestinal, for example: esophagus (squamous cell        carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma), stomach        (carcinoma, lymphoma, leiomyosarcoma), pancreas (ductal        adenocarcinoma, insulinoma, glucagonoma, gastrinoma, carcinoid        tumors, vipoma), small bowel (adenocarcinoma, lymphoma,        carcinoid tumors, Kaposi's sarcoma, leiomyoma, hemangioma,        lipoma, neurofibroma, fibroma), large bowel (adenocarcinoma,        tubular adenoma, villous adenoma, hamartoma, leiomyoma);        Genitourinary tract, for example: kidney (adenocarcinoma, Wilm's        tumor (nephroblastoma), lymphoma, leukemia), bladder and urethra        (squamous cell carcinoma, transitional cell carcinoma,        adenocarcinoma), prostate (adenocarcinoma, sarcoma), testis        (seminoma, teratoma, embryonal carcinoma, teratocarcinoma,        choriocarcinoma, sarcoma, interstitial cell carcinoma, fibroma,        fibroadenoma, adenomatoid tumors, lipoma);    -   Liver, for example: hepatoma (hepatocellular carcinoma),        choiangiocarcinoma, hepatoblastoma, angiosarcoma, hepatocellular        adenoma, hemangioma;    -   Biliary tract, for example: gall bladder carcinoma, ampullary        carcinoma, choiangiocarcinoma;    -   Bone, for example: osteogenic sarcoma (osteosarcoma),        fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma,        Ewing's sarcoma, malignant lymphoma (reticulum cell sarcoma),        multiple myeloma, malignant giant cell tumor chordoma,        osteochronfroma (osteocartilaginous exostoses), benign        chondroma, chondroblastoma, chondromyxofibroma, osteoid osteoma        and giant cell tumors;    -   Nervous system, for example: skull (osteoma, hemangioma,        granuloma, xanthoma, osteitis deformans), meninges (meningioma,        meningiosarcoma, gliomatosis), brain (astrocytoma,        medulloblastoma, glioma, ependymoma, germinoma (pinealoma),        glioblastoma multiform, oligodendroglioma, schwannoma,        retinoblastoma, congenital tumors), spinal cord neurofibroma,        neurofibromatosis type 1, meningioma, glioma, sarcoma);    -   Gynecological, for example: uterus (endometrial carcinoma,        uterine carcinoma, uterine corpus endometrial carcinoma), cervix        (cervical carcinoma, pre-tumor cervical dysplasia), ovaries        (ovarian carcinoma (serous cystadenocarcinoma, mucinous        cystadenocarcinoma, unclassified carcinoma), granulosa-thecal        cell tumors, Sertoli-Leydig cell tumors, dysgerminoma, malignant        teratoma), vulva (squamous ceil carcinoma, intraepithelial        carcinoma, adenocarcinoma, fibrosarcoma, melanoma), vagina        (clear cell carcinoma, squamous cell carcinoma, botryoid sarcoma        (embryonal rhabdomyosarcoma), fallopian tubes (carcinoma);    -   Hematologic, for example: blood (myeloid leukemia (acute and        chronic), acute lymphoblastic leukemia, chronic lymphocytic        leukemia, myeloproliferative diseases (e.g., myelofibrosis and        myeloproliferative neoplasms, multiple myeloma, myelodysplastic        syndrome), Hodgkin's disease, non-Hodgkin's lymphoma (malignant        lymphoma);    -   Skin, for example: malignant melanoma, basal cell carcinoma,        squamous ceil carcinoma, Kaposi's sarcoma, moles dysplastic        nevi, lipoma, angioma, dermatofibroma, keloids, psoriasis; and    -   Adrenal glands, for example: neuroblastoma.

In some embodiments, the Ras protein is wild-type. (Ras^(WT)).Accordingly, in some embodiments, a compound of the present invention isemployed in a method of treating a patient having a cancer comprising aRas^(WT) (e.g., K-Ras^(WT), H-Ras^(WT) or N-Ras^(WT)). In someembodiments, the Ras protein is Ras amplification (e.g., K-Ras^(amp)).Accordingly, in some embodiments, a compound of the present invention isemployed in a method of treating a patient having a cancer comprising aRas^(amp) (K-Ras^(amp), H-Ras^(amp) or N-Ras^(amp)). In someembodiments, the cancer comprises a Ras mutation, such as a Ras mutationdescribed herein. In some embodiments, a mutation is selected from:

-   -   (a) the following K-Ras mutants: G12D, G12V, G12C, G13D, G12R,        G12A, Q61H, G12S, A146T, G13C, Q61L, Q61R, K117N, A146V, G12F,        Q61K, L19F, Q22K, V14I, A59T, A146P, G13R, G12L, or G13V, and        combinations thereof;    -   (b) the following H-Ras mutants: Q61R, G13R, Q61K, G12S, Q61L,        G12D, G13V, G13D, G12C, K117N, A59T, G12V, G13C, Q61H, G13S,        A18V, D119N, G13N, A146T, A66T, G12A, A146V, G12N, or G12R, and        combinations thereof; and    -   (c) the following N-Ras mutants: Q61R, Q61K, G12D, Q61L, Q61H,        G13R, G13D, G12S, G12C, G12V, G12A, G13V, G12R, P185S, G13C,        A146T, G60E, Q61P, A59D, E132K, E49K, T50I, A146V, or A59T, and        combinations thereof;        or a combination of any of the foregoing. In some embodiments,        the cancer comprises a Ras mutation selected from the group        consisting of G12C, G13C, G12A, G12D, G13D, G12S, G13S, G12V and        G13V. In some embodiments, the cancer comprises at least two Ras        mutations selected from the group consisting of G12C, G13C,        G12A, G12D, G13D, G12S, G13S, G12V and G13V. In some        embodiments, a compound of the present invention inhibits more        than one Ras mutant. For example, a compound may inhibit both        K-Ras G12D and K-Ras G12C. In some embodiments, a compound may        inhibit both K-Ras G12V and K-Ras G12C. In some embodiments, a        compound may inhibit both K-Ras G12C and K-Ras G13C. In some        embodiments, a compound may inhibit both K-Ras G12D and K-Ras        G12V. In some embodiments, a compound may inhibit both K-Ras        G12V and K-Ras G12S. In some embodiments, the mutation is        selected from the group consisting of G12A, G12C, G12D, G12E,        G12F, G12H, G12I, G12K, G12L, G12M, G12N, G12P, G12Q, G12R,        G12S, G12T, G12V, G12W and G12Y, or a combination thereof, of        K-Ras, N-Ras or H-Ras. In some embodiments, the mutation is        selected from the group consisting of G12H, G12I, G12K, G12M,        G12N, G12P, G12Q, G12T, G12W, and G12Y, or a combination        thereof, of K-Ras, N-Ras or H-Ras. In some embodiments, the        compound inhibits wild-type K-Ras, wild-type H-Ras or wild-type        N-Ras, and optionally further inhibits a mutated Ras protein        containing a mutation as described herein. In some embodiments,        the cancer is non-small cell lung cancer and the Ras mutation        comprises a K-Ras mutation, such as K-Ras G12C. In some        embodiments, the cancer is colorectal cancer and the Ras        mutation comprises a K-Ras mutation, such as K-Ras G12C. In some        embodiments, the cancer is pancreatic cancer and the Ras        mutation comprises an N-Ras mutation, such as N-Ras G12D. In        some embodiments, the cancer is non-small cell lung cancer and        the Ras protein is K-Ras^(amp).

Additionally, in some embodiments, the cancer comprises a K-Ras mutationselected from the group consisting of G12C, G12D, G13C, G12V, G13D,G12R, G12S, Q61H, Q61K and Q61L. In some embodiments, the cancercomprises an N-Ras mutation selected from the group consisting of G12C,Q61H, Q61K, Q61L, Q61P and Q61R. In some embodiments, the cancercomprises an H-Ras mutation selected from the group consisting of Q61Hand Q61L. In some embodiments, the cancer comprises a Ras mutationselected from the group consisting of G12C, G13C, G12A, G12D, G13D,G12S, G13S, G12V and G13V. In some embodiments, the cancer comprises atleast two Ras mutations selected from the group consisting of G12C,G13C, G12A, G12D, G13D, G12S, G13S, G12V and G13V. In some embodiments,a compound of the present invention inhibits more than one Ras mutant.For example, a compound may inhibit both K-Ras G12C and K-Ras G13C. Acompound may inhibit both N-Ras G12C and K-Ras G12C. In someembodiments, a compound may inhibit both K-Ras G12C and K-Ras G12D. Insome embodiments, a compound may inhibit both K-Ras G12V and K-Ras G12C.In some embodiments, a compound may inhibit both K-Ras G12V and K-RasG12S. In some embodiments, a compound of the present invention inhibitsRas^(wt) in addition to one or more additional Ras mutations (e.g., K-,H- or N-Ras^(wt) and K-Ras G12D, G12V, G12C, G13D, G12R, G12A, Q61H,G12S, A146T, G13C, Q61L, Q61R, K117N, A146V, G12F, Q61K, L19F, Q22K,V14I, A59T, A146P, G13R, G12L, or G13V; K-, H- or N-Ras^(wt) and H-RasQ61R, G13R, Q61K, G12S, Q61L, G12D, G13V, G13D, G12C, K117N, A59T, G12V,G13C, Q61H, G13S, A18V, D119N, G13N, A146T, A66T, G12A, A146V, G12N, orG12R; or K-, H- or N-Ras^(wt) and N-Ras Q61R, Q61K, G12D, Q61L, Q61H,G13R, G13D, G12S, G12C, G12V, G12A, G13V, G12R, P185S, G13C, A146T,G60E, Q61P, A59D, E132K, E49K, T50I, A146V, or A59T). In someembodiments, a compound of the present invention inhibits Ras^(amp) inaddition to one or more additional Ras mutations (e.g., K-, H- orN-Ras^(amp) and K-Ras G12D, G12V, G12C, G13D, G12R, G12A, Q61H, G12S,A146T, G13C, Q61L, Q61R, K117N, A146V, G12F, Q61K, L19F, Q22K, V14I,A59T, A146P, G13R, G12L, or G13V; K-, H- or N-Ras^(amp) and H-Ras Q61R,G13R, Q61K, G12S, Q61L, G12D, G13V, G13D, G12C, K117N, A59T, G12V, G13C,Q61H, G13S, A18V, D119N, G13N, A146T, A66T, G12A, A146V, G12N, or G12R;or K-, H- or N-Ras^(amp) and N-Ras Q61R, Q61K, G12D, Q61L, Q61H, G13R,G13D, G12S, G12C, G12V, G12A, G13V, G12R, P185S, G13C, A146T, G60E,Q61P, A59D, E132K, E49K, T50I, A146V, or A59T).

Methods of detecting Ras mutations are known in the art. Such meansinclude, but are not limited to direct sequencing, and utilization of ahigh-sensitivity diagnostic assay (with CE-IVD mark), e.g., as describedin Domagala, et al., Pol J Pathol 3: 145-164 (2012), incorporated hereinby reference in its entirety, including TheraScreen PCR; AmoyDx;PNACIamp; RealQuality; EntroGen; LightMix; StripAssay; Hybcell plexA;Devyser; Surveyor; Cobas; and TheraScreen Pyro. See, also, e.g., WO2020/106640.

In some embodiments, the cancer is non-small cell lung cancer and theRas mutation comprises a K-Ras mutation, such as K-Ras G12C, K-Ras G12Vor K-Ras G12D. In some embodiments, the cancer is colorectal cancer andthe Ras mutation comprises a K-Ras mutation, such as K-Ras G12C, K-RasG12V or K-Ras G12D. In some embodiments, the cancer is pancreatic cancerand the Ras mutation comprises an K-Ras mutation, such as K-Ras G12D orK-Ras G12V. In some embodiments, the cancer is pancreatic cancer and theRas mutation comprises an N-Ras mutation, such as N-Ras G12D. In someembodiments, the cancer is melanoma and the Ras mutation comprises anN-Ras mutation, such as N-Ras Q61R or N-Ras Q61K. In some embodiments,the cancer is non-small cell lung cancer and the Ras protein isK-Ras^(amp). In any of the foregoing if not already specified, acompound may inhibit Ras^(WT) (e.g., K-, H- or N-Ras^(WT)) or Ras^(amp)(e.g., K-, H- or N-Ras^(amp)) as well.

In some embodiments, a cancer comprises a Ras mutation and anSTK11^(LOF), a KEAP1, an EPHA5 or an NF1 mutation, or a combinationthereof. In some embodiments, the cancer is non-small cell lung cancerand comprises a K-Ras G12C mutation. In some embodiments, the cancer isnon-small cell lung cancer and comprises a K-Ras G12C mutation, anSTK11^(LOF) mutation, and a KEAP1 mutation. In some embodiments, thecancer is non-small cell lung cancer and comprises a K-Ras G12C mutationand an STK11^(LOF) mutation. In some embodiments, the cancer isnon-small cell lung cancer and comprises a K-Ras G12C mutation and anSTK11^(LOF) mutation. In some embodiments, a cancer comprises a K-RasG13C Ras mutation and an STK11^(LOF), a KEAP1, an EPHA5 or an NF1mutation. In some embodiments, the cancer is non-small cell lung cancerand comprises a K-Ras G12D mutation. In some embodiments, the cancer isnon-small cell lung cancer and comprises a K-Ras G12V mutation. In someembodiments, the cancer is colorectal cancer and comprises a K-Ras G12Cmutation. In some embodiments, the cancer is pancreatic cancer andcomprises a K-Ras G12D mutation. In some embodiments, the cancer ispancreatic cancer and comprises a K-Ras G12V mutation. In someembodiments, the cancer is endometrial cancer and comprises a K-Ras G12Cmutation. In some embodiments, the cancer is gastric cancer andcomprises a K-Ras G12C mutation. In any of the foregoing, a compound mayinhibit Ras^(wt) (e.g., K-, H- or N-Ras^(wt)) or Ras^(amp) (e.g., K-, H-or N-Ras^(amp)) as well.

Also provided is a method of inhibiting a Ras protein in a cell, themethod comprising contacting the cell with an effective amount of acompound of the present invention, or a pharmaceutically acceptable saltthereof. A compound, or a pharmaceutically acceptable salt thereof, mayinhibit more than one type of Ras protein in a cell. A method ofinhibiting RAF-Ras binding, the method comprising contacting the cellwith an effective amount of a compound of the present invention, or apharmaceutically acceptable salt thereof, is also provided. The cell maybe a cancer cell. The cancer cell may be of any type of cancer describedherein. The cell may be in vivo or in vitro.

Combination Therapy

The methods of the invention may include a compound of the inventionused alone or in combination with one or more additional therapies(e.g., non-drug treatments or therapeutic agents). The dosages of one ormore of the additional therapies (e.g., non-drug treatments ortherapeutic agents) may be reduced from standard dosages whenadministered alone. For example, doses may be determined empiricallyfrom drug combinations and permutations or may be deduced byisobolographic analysis (e.g., Black et al., Neurology 65:S3-S6 (2005)).

A compound of the present invention may be administered before, after,or concurrently with one or more of such additional therapies. Whencombined, dosages of a compound of the invention and dosages of the oneor more additional therapies (e.g., non-drug treatment or therapeuticagent) provide a therapeutic effect (e.g., synergistic or additivetherapeutic effect). A compound of the present invention and anadditional therapy, such as an anti-cancer agent, may be administeredtogether, such as in a unitary pharmaceutical composition, or separatelyand, when administered separately, this may occur simultaneously orsequentially. Such sequential administration may be close or remote intime.

In some embodiments, the additional therapy is the administration ofside-effect limiting agents (e.g., agents intended to lessen theoccurrence or severity of side effects of treatment. For example, insome embodiments, the compounds of the present invention can also beused in combination with a therapeutic agent that treats nausea.Examples of agents that can be used to treat nausea include: dronabinol,granisetron, metoclopramide, ondansetron, and prochlorperazine, orpharmaceutically acceptable salts thereof.

In some embodiments, the one or more additional therapies includes anon-drug treatment (e.g., surgery or radiation therapy). In someembodiments, the one or more additional therapies includes a therapeuticagent (e.g., a compound or biologic that is an anti-angiogenic agent,signal transduction inhibitor, antiproliferative agent, glycolysisinhibitor, or autophagy inhibitor). In some embodiments, the one or moreadditional therapies includes a non-drug treatment (e.g., surgery orradiation therapy) and a therapeutic agent (e.g., a compound or biologicthat is an anti-angiogenic agent, signal transduction inhibitor,antiproliferative agent, glycolysis inhibitor, or autophagy inhibitor).In other embodiments, the one or more additional therapies includes twotherapeutic agents. In still other embodiments, the one or moreadditional therapies includes three therapeutic agents. In someembodiments, the one or more additional therapies includes four or moretherapeutic agents.

In this Combination Therapy section, all references are incorporated byreference for the agents described, or a pharmaceutically acceptablesalt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomerthereof, whether explicitly stated as such or not.

Non-Drug Therapies

Examples of non-drug treatments include, but are not limited to,radiation therapy, cryotherapy, hyperthermia, surgery (e.g., surgicalexcision of tumor tissue), and T cell adoptive transfer (ACT) therapy.

In some embodiments, the compounds of the invention may be used as anadjuvant therapy after surgery. In some embodiments, the compounds ofthe invention may be used as a neo-adjuvant therapy prior to surgery.

Radiation therapy may be used for inhibiting abnormal cell growth ortreating a hyperproliferative disorder, such as cancer, in a subject(e.g., mammal (e.g., human)). Techniques for administering radiationtherapy are known in the art. Radiation therapy can be administeredthrough one of several methods, or a combination of methods, including,without limitation, external-beam therapy, internal radiation therapy,implant radiation, stereotactic radiosurgery, systemic radiationtherapy, radiotherapy and permanent or temporary interstitial brachytherapy. The term “brachy therapy,” as used herein, refers to radiationtherapy delivered by a spatially confined radioactive material insertedinto the body at or near a tumor or other proliferative tissue diseasesite. The term is intended, without limitation, to include exposure toradioactive isotopes (e.g., At-211, I-131, I-125, Y-90, Re-186, Re-188,Sm-153, Bi-212, P-32, and radioactive isotopes of Lu). Suitableradiation sources for use as a cell conditioner of the present inventioninclude both solids and liquids. By way of non-limiting example, theradiation source can be a radionuclide, such as I-125, I-131, Yb-169,Ir-192 as a solid source, I-125 as a solid source, or otherradionuclides that emit photons, beta particles, gamma radiation, orother therapeutic rays. The radioactive material can also be a fluidmade from any solution of radionuclide(s), e.g., a solution of I-125 orI-131, or a radioactive fluid can be produced using a slurry of asuitable fluid containing small particles of solid radionuclides, suchas Au-198, or Y-90. Moreover, the radionuclide(s) can be embodied in agel or radioactive micro spheres.

In some embodiments, the compounds of the present invention can renderabnormal cells more sensitive to treatment with radiation for purposesof killing or inhibiting the growth of such cells. Accordingly, thisinvention further relates to a method for sensitizing abnormal cells ina mammal to treatment with radiation which comprises administering tothe mammal an amount of a compound of the present invention, whichamount is effective to sensitize abnormal cells to treatment withradiation. The amount of the compound in this method can be determinedaccording to the means for ascertaining effective amounts of suchcompounds described herein. In some embodiments, the compounds of thepresent invention may be used as an adjuvant therapy after radiationtherapy or as a neo-adjuvant therapy prior to radiation therapy.

In some embodiments, the non-drug treatment is a T cell adoptivetransfer (ACT) therapy. In some embodiments, the T cell is an activatedT cell. The T cell may be modified to express a chimeric antigenreceptor (CAR). CAR modified T (CAR-T) cells can be generated by anymethod known in the art. For example, the CAR-T cells can be generatedby introducing a suitable expression vector encoding the CAR to a Tcell. Prior to expansion and genetic modification of the T cells, asource of T cells is obtained from a subject. T cells can be obtainedfrom a number of sources, including peripheral blood mononuclear cells,bone marrow, lymph node tissue, cord blood, thymus tissue, tissue from asite of infection, ascites, pleural effusion, spleen tissue, and tumors.In certain embodiments of the present invention, any number of T celllines available in the art may be used. In some embodiments, the T cellis an autologous T cell. Whether prior to or after genetic modificationof the T cells to express a desirable protein (e.g., a CAR), the T cellscan be activated and expanded generally using methods as described, forexample, in U.S. Pat. Nos. 6,352,694; 6,534,055; 6,905,680; 6,692,964;5,858,358; 6,887,466; 6,905,681; 7,144,575; 7,067,318; 7,172,869;7,232,566; 7,175,843; 7,572,631; 5,883,223; 6,905,874; 6,797,514; and6,867,041.

Therapeutic Agents

A therapeutic agent may be a compound used in the treatment of cancer orsymptoms associated therewith.

For example, a therapeutic agent may be a steroid. Accordingly, in someembodiments, the one or more additional therapies includes a steroid.Suitable steroids may include, but are not limited to,21-acetoxypregnenolone, alclometasone, algestone, amcinonide,beclomethasone, betamethasone, budesonide, chloroprednisone, clobetasol,clocortolone, cloprednol, corticosterone, cortisone, cortivazol,deflazacort, desonide, desoximetasone, dexamethasone, diflorasone,diflucortolone, difuprednate, enoxolone, fluazacort, flucloronide,flumethasone, flunisolide, fluocinolone acetonide, fluocinonide,fluocortin butyl, fluocortolone, fluorometholone, fluperolone acetate,fluprednidene acetate, fluprednisolone, flurandrenolide, fluticasonepropionate, formocortal, halcinonide, halobetasol propionate,halometasone, hydrocortisone, loteprednol etabonate, mazipredone,medrysone, meprednisone, methylprednisolone, mometasone furoate,paramethasone, prednicarbate, prednisolone, prednisolone25-diethylaminoacetate, prednisolone sodium phosphate, prednisone,prednival, prednylidene, rimexolone, tixocortol, triamcinolone,triamcinolone acetonide, triamcinolone benetonide, triamcinolonehexacetonide, and salts or derivatives thereof.

Further examples of therapeutic agents that may be used in combinationtherapy with a compound of the present invention include compoundsdescribed in the following patents: U.S. Pat. Nos. 6,258,812, 6,630,500,6,515,004, 6,713,485, 5,521,184, 5,770,599, 5,747,498, 5,990,141,6,235,764, and 8,623,885, and International Patent ApplicationsWO01/37820, WO01/32651, WO02/68406, WO02/66470, WO02/55501, WO04/05279,WO04/07481, WO04/07458, WO04/09784, WO02/59110, WO99/45009, WO00/59509,WO99/61422, WO00/12089, and WO00/02871.

A therapeutic agent may be a biologic (e.g., cytokine (e.g., interferonor an interleukin such as IL-2)) used in treatment of cancer or symptomsassociated therewith. In some embodiments, the biologic is animmunoglobulin-based biologic, e.g., a monoclonal antibody (e.g., ahumanized antibody, a fully human antibody, an Fc fusion protein, or afunctional fragment thereof) that agonizes a target to stimulate ananti-cancer response or antagonizes an antigen important for cancer.Also included are antibody-drug conjugates.

A therapeutic agent may be a T-cell checkpoint inhibitor. In oneembodiment, the checkpoint inhibitor is an inhibitory antibody (e.g., amonospecific antibody such as a monoclonal antibody). The antibody maybe, e.g., humanized or fully human. In some embodiments, the checkpointinhibitor is a fusion protein, e.g., an Fc-receptor fusion protein. Insome embodiments, the checkpoint inhibitor is an agent, such as anantibody, that interacts with a checkpoint protein. In some embodiments,the checkpoint inhibitor is an agent, such as an antibody, thatinteracts with the ligand of a checkpoint protein. In some embodiments,the checkpoint inhibitor is an inhibitor (e.g., an inhibitory antibodyor small molecule inhibitor) of CTLA-4 (e.g., an anti-CTLA-4 antibody orfusion a protein). In some embodiments, the checkpoint inhibitor is aninhibitor or antagonist (e.g., an inhibitory antibody or small moleculeinhibitor) of PD-1. In some embodiments, the checkpoint inhibitor is aninhibitor or antagonist (e.g., an inhibitory antibody or small moleculeinhibitor) of PD-L1. In some embodiments, the checkpoint inhibitor is aninhibitor or antagonist (e.g., an inhibitory antibody or Fc fusion orsmall molecule inhibitor) of PD-L2 (e.g., a PD-L2/lg fusion protein). Insome embodiments, the checkpoint inhibitor is an inhibitor or antagonist(e.g., an inhibitory antibody or small molecule inhibitor) of B7-H3,B7-H4, BTLA, HVEM, TIM3, GAL9, LAG3, VISTA, KIR, 2B4, CD160, CGEN-15049,CHK 1, CHK2, A2aR, B-7 family ligands, or a combination thereof. In someembodiments, the checkpoint inhibitor is pembrolizumab, nivolumab,PDR001 (NVS), REGN2810 (Sanofi/Regeneron), a PD-L1 antibody such as,e.g., avelumab, durvalumab, atezolizumab, pidilizumab, JNJ-63723283(JNJ), BGB-A317 (BeiGene & Celgene) or a checkpoint inhibitor disclosedin Preusser, M. et al. (2015) Nat. Rev. Neurol., including, withoutlimitation, ipilimumab, tremelimumab, nivolumab, pembrolizumab, AMP224,AMP514/MEDI0680, BMS936559, MEDI4736, MPDL3280A, MSB0010718C, BMS986016,IMP321, lirilumab, IPH2101, 1-7F9, and KW-6002.

A therapeutic agent may be an anti-TIGIT antibody, such as MBSA43,BMS-986207, MK-7684, COM902, AB154, MTIG7192A or OMP-313M32(etigilimab).

A therapeutic agent may be an agent that treats cancer or symptomsassociated therewith (e.g., a cytotoxic agent, non-peptide smallmolecules, or other compound useful in the treatment of cancer orsymptoms associated therewith, collectively, an “anti-cancer agent”).Anti-cancer agents can be, e.g., chemotherapeutics or targeted therapyagents.

Anti-cancer agents include mitotic inhibitors, intercalatingantibiotics, growth factor inhibitors, cell cycle inhibitors, enzymes,topoisomerase inhibitors, biological response modifiers, alkylatingagents, antimetabolites, folic acid analogs, pyrimidine analogs, purineanalogs and related inhibitors, vinca alkaloids, epipodopyyllotoxins,antibiotics, L-Asparaginase, topoisomerase inhibitors, interferons,platinum coordination complexes, anthracenedione substituted urea,methyl hydrazine derivatives, adrenocortical suppressant,adrenocorticosteroides, progestins, estrogens, antiestrogen, androgens,antiandrogen, and gonadotropin-releasing hormone analog. Furtheranti-cancer agents include leucovorin (LV), irenotecan, oxaliplatin,capecitabine, paclitaxel, and doxetaxel. In some embodiments, the one ormore additional therapies includes two or more anti-cancer agents. Thetwo or more anti-cancer agents can be used in a cocktail to beadministered in combination or administered separately. Suitable dosingregimens of combination anti-cancer agents are known in the art anddescribed in, for example, Saltz et al., Proc. Am. Soc. Clin. Oncol.18:233a (1999), and Douillard et al., Lancet 355(9209):1041-1047 (2000).

Other non-limiting examples of anti-cancer agents include Gieevec®(Imatinib Mesylate); Kyproiis® (carfilzomib); Velcade® (bortezomib);Casodex (bicalutamide); Iressa® (gefitinib); alkylating agents such asthiotepa and cyclosphosphamide; alkyl sulfonates such as busulfan,improsulfan and piposulfan; aziridines such as benzodopa, carboquone,meturedopa, and uredopa; ethylenimines and methylamelamines includingaltretamine, triethylenemelamine, triethylenephosphoramide,triethiylenethiophosphoramide and trimethylolomelamine; acetogenins(especially bullatacin and bullatacinone); a camptothecin (including thesynthetic analogue topotecan); bryostatin; callystatin; CC-1065(including its adozelesin, carzelesin and bizelesin syntheticanalogues); cryptophycins (particularly cryptophycin 1 and cryptophycin8); dolastatin; duocarmycin (including the synthetic analogues, KW-2189and CB1-TM1); eleutherobin; pancratistatin; sarcodictyin A;spongistatin; nitrogen mustards such as chlorambucil, chlornaphazine,cholophosphamide, estramustine, ifosfamide, mechlorethamine,mechlorethamine oxide hydrochloride, melphalan, novembichin,phenesterine, prednimustine, trofosfamide, uracil mustard; nitrosureassuch as carmustine, chlorozotocin, fotemustine, lomustine, nimustine,and ranimustine; antibiotics such as the enediyne antibiotics (e.g.,calicheamicin, such as calicheamicin gammall and calicheamicin omegall(see, e.g., Agnew, Chem. Inti. Ed Engl. 33:183-186 (1994)); dynemicinsuch as dynemicin A; bisphosphonates such as clodronate; an esperamicin;neocarzinostatin chromophore and related chromoprotein enediyneantiobiotic chromophores, aclacinomysins, actinomycin, authramycin,azaserine, bleomycins, cactinomycin, calicheamicin, carabicin,caminomycin, carminomycin, carzinophilin, chromomycins, dactinomycin,daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, adriamycin(doxorubicin), morpholino-doxorubicin, cyanomorpholino-doxorubicin,2-pyrrolino-doxorubicin, deoxydoxorubicin, epirubicin, esorubicin,idarubicin, marcellomycin, mitomycins such as mitomycin C, mycophenolicacid, nogalamycin, olivomycins, peplomycin, potfiromycin, puromycin,quelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin,ubenimex, zinostatin, zorubicin; anti-metabolites such as methotrexateand 5-fluorouracil (5-FU); folic acid analogues such as denopterin,pteropterin, trimetrexate; purine analogs such as fludarabine,6-mercaptopurine, thiamiprine, thioguanine; pyrimidine analogs such asancitabine, azacitidine, 6-azauridine, carmofur, cytarabine,dideoxyuridine, doxifluridine, enocitabine, floxuridine; androgens suchas calusterone, dromostanolone propionate, epitiostanol, mepitiostane,testolactone; anti-adrenals such as aminoglutethimide, mitotane,trilostane; folic acid replenishers such as frolinic acid; aceglatone;aldophosphamide glycoside; aminolevulinic acid; eniluracil; amsacrine;bestrabucil; bisantrene; edatraxate; defofamine; demecolcine;diaziquone; elfomithine; elliptinium acetate; an epothilone such asepothilone B; etoglucid; gallium nitrate; hydroxyurea; lentinan;lonidamine; maytansinoids such as maytansine and ansamitocins;mitoguazone; mitoxantrone; mopidamol; nitracrine; pentostatin; phenamet;pirarubicin; losoxantrone; podophyllinic acid; 2-ethylhydrazide;procarbazine; PSK® polysaccharide complex (JHS Natural Products, Eugene,Oreg.); razoxane; rhizoxin; sizofiran; spirogermanium; tenuazonic acid;triaziquone; 2,2′,2″-trichlorotriethylamine; trichothecenes such as T-2toxin, verracurin A, roridin A and anguidine; urethane; vindesine;dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman;gacytosine; arabinoside (“Ara-C”); cyclophosphamide; thiotepa; taxoids,e.g., Taxol® (paclitaxel), Abraxane® (cremophor-free, albumin-engineerednanoparticle formulation of paclitaxel), and Taxotere® (doxetaxel);chloranbucil; tamoxifen (Nolvadex™); raloxifene; aromatase inhibiting4(5)-imidazoles; 4-hydroxyiamoxifen; trioxifene; keoxifene; LY 117018;onapristone; toremifene (Fareston®); flutamide, nilutamide,bicalutamide, leuproiide, gosereiin; chlorambucil; Gemzar® gemcitabine;6-thioguanine; mercaptopurine; platinum coordination complexes such ascisplatin, oxaliplatin and carboplatin; vinblastine; platinum; etoposide(VP-16); ifosfamide; mitoxantrone; vincristine; Navelbine®(vinorelbine); novantrone; teniposide; edatrexate; daunomycin;aminopterin; ibandronate; irinotecan (e.g., CPT-11); topoisomeraseinhibitor RFS 2000; difluoromethylornithine (DMFO); retinoids such asretinoic acid; esperamicins; capecitabine (e.g., Xeloda®); andpharmaceutically acceptable salts of any of the above.

Additional non-limiting examples of anti-cancer agents includetrastuzumab (Herceptin®), bevacizumab (Avastin®), cetuximab (Erbitux®),rituximab (Rituxan®), Taxol®, Arimidex®, ABVD, avicine, abagovomab,acridine carboxamide, adecatumumab,17-N-allylamino-17-demethoxygeldanamycin, alpharadin, alvocidib,3-aminopyridine-2-carboxaldehyde thiosemicarbazone, amonafide,anthracenedione, anti-CD22 immunotoxins, antineoplastics (e.g.,cell-cycle nonspecific antineoplastic agents, and other antineoplasticsdescribed herein), antitumorigenic herbs, apaziquone, atiprimod,azathioprine, belotecan, bendamustine, BIBW2992, biricodar,brostallicin, bryostatin, buthionine sulfoximine, CBV (chemotherapy),calyculin, dichloroacetic acid, discodermolide, elsamitrucin,enocitabine, eribulin, exatecan, exisulind, ferruginol, forodesine,fosfestrol, ICE chemotherapy regimen, IT-101, imexon, imiquimod,indolocarbazole, irofulven, laniquidar, larotaxel, lenalidomide,lucanthone, lurtotecan, mafosfamide, mitozolomide, nafoxidine,nedaplatin, olaparib, ortataxel, PAC-1, pawpaw, pixantrone, proteasomeinhibitors, rebeccamycin, resiquimod, rubitecan, SN-38, salinosporamideA, sapacitabine, Stanford V, swainsonine, talaporfin, tariquidar,tegafur-uracil, temodar, tesetaxel, triplatin tetranitrate,tris(2-chloroethyl)amine, troxacitabine, uramustine, vadimezan,vinflunine, ZD6126, and zosuquidar.

Further non-limiting examples of anti-cancer agents include naturalproducts such as vinca alkaloids (e.g., vinblastine, vincristine, andvinorelbine), epidipodophyllotoxins (e.g., etoposide and teniposide),antibiotics (e.g., dactinomycin (actinomycin D), daunorubicin, andidarubicin), anthracyclines, mitoxantrone, bleomycins, plicamycin(mithramycin), mitomycin, enzymes (e.g., L-asparaginase whichsystemically metabolizes L-asparagine and deprives cells which do nothave the capacity to synthesize their own asparagine), antiplateletagents, antiproliferative/antimitotic alkylating agents such as nitrogenmustards (e.g., mechlorethamine, cyclophosphamide and analogs,melphalan, and chlorambucil), ethylenimines and methylmelamines (e.g.,hexaamethylmelaamine and thiotepa), CDK inhibitors (e.g., a CDK4/6inhibitor such as abemaciclib, ribociclib, palbociclib; seliciclib,UCN-01, P1446A-05, PD-0332991, dinaciclib, P27-00, AT-7519, RGB286638,and SCH727965), alkyl sulfonates (e.g., busulfan), nitrosoureas (e.g.,carmustine (BCNU) and analogs, and streptozocin), trazenes-dacarbazinine(DTIC), antiproliferative/antimitotic antimetabolites such as folic acidanalogs, pyrimidine analogs (e.g., fluorouracil, floxuridine, andcytarabine), purine analogs and related inhibitors (e.g.,mercaptopurine, thioguanine, pentostatin, and 2-chlorodeoxyadenosine),aromatase inhibitors (e.g., anastrozole, exemestane, and letrozole), andplatinum coordination complexes (e.g., cisplatin and carboplatin),procarbazine, hydroxyurea, mitotane, aminoglutethimide, histonedeacetylase (HDAC) inhibitors (e.g., trichostatin, sodium butyrate,apicidan, suberoyl anilide hydroamic acid, vorinostat, LBH 589,romidepsin, ACY-1215, and panobinostat), mTOR inhibitors (e.g.,vistusertib, temsirolimus, everolimus, ridaforolimus, and sirolimus),KSP(Eg5) inhibitors (e.g., Array 520), DNA binding agents (e.g.,Zalypsis®), PI3K inhibitors such as PI3K delta inhibitor (e.g., GS-1101and TGR-1202), PI3K delta and gamma inhibitor (e.g., CAL-130),copanlisib, alpelisib and idelalisib; multi-kinase inhibitor (e.g., TG02and sorafenib), hormones (e.g., estrogen) and hormone agonists such asleutinizing hormone releasing hormone (LHRH) agonists (e.g., goserelin,leuprolide and triptorelin), BAFF-neutralizing antibody (e.g.,LY2127399), IKK inhibitors, p38MAPK inhibitors, anti-IL-6 (e.g.,CNT0328), telomerase inhibitors (e.g., GRN 163L), aurora kinaseinhibitors (e.g., MLN8237), cell surface monoclonal antibodies (e.g.,anti-CD38 (HUMAX-CD38), anti-CSI (e.g., elotuzumab), HSP90 inhibitors(e.g., 17 AAG and KOS 953), P13K/Akt inhibitors (e.g., perifosine), Aktinhibitors (e.g., GSK-2141795), PKC inhibitors (e.g., enzastaurin), FTIs(e.g., Zarnestra™), anti-CD138 (e.g., BT062), Torcl/2 specific kinaseinhibitors (e.g., INK128), ER/UPR targeting agents (e.g., MKC-3946),cFMS inhibitors (e.g., ARRY-382), JAK1/2 inhibitors (e.g., CYT387), PARPinhibitors (e.g., olaparib and veliparib (ABT-888)), and BCL-2antagonists.

In some embodiments, an anti-cancer agent is selected frommechlorethamine, camptothecin, ifosfamide, tamoxifen, raloxifene,gemcitabine, Navelbine®, sorafenib, or any analog or derivative variantof the foregoing.

In some embodiments, the anti-cancer agent is a HER2 inhibitor.Non-limiting examples of HER2 inhibitors include monoclonal antibodiessuch as trastuzumab (Herceptin®) and pertuzumab (Perjeta®); smallmolecule tyrosine kinase inhibitors such as gefitinib (Iressa®),erlotinib (Tarceva®), pilitinib, CP-654577, CP-724714, canertinib (CI1033), HKI-272, lapatinib (GW-572016; Tykerb®), PKI-166, AEE788,BMS-599626, HKI-357, BIBW 2992, ARRY-334543, and JNJ-26483327.

In some embodiments, an anti-cancer agent is an ALK inhibitor.Non-limiting examples of ALK inhibitors include ceritinib, TAE-684(NVP-TAE694), PF02341066 (crizotinib or 1066), alectinib; brigatinib;entrectinib; ensartinib (X-396); lorlatinib; ASP3026; CEP-37440;4SC-203; TL-398; PLB1003; TSR-011; CT-707; TPX-0005, and AP26113.Additional examples of ALK kinase inhibitors are described in examples3-39 of WO 05016894.

In some embodiments, an anti-cancer agent is an inhibitor of a memberdownstream of a Receptor Tyrosine Kinase (RTK)/Growth Factor Receptor(e.g., a SHP2 inhibitor (e.g., SHP099, TNO155, RMC-4550, RMC-4630,JAB-3068, JAB-3312, RLY-1971, ERAS-601, SH3809, PF-07284892, or BBP-398,or a pharmaceutically acceptable salt, solvate, isomer (e.g.,stereoisomer), prodrug, or tautomer thereof), a SOS1 inhibitor (e.g.,BI-1701963, BI-3406, SDR5, BAY-293, or RMC-5845, or a pharmaceuticallyacceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, ortautomer thereof), a Raf inhibitor, a MEK inhibitor, an ERK inhibitor, aPI3K inhibitor, a PTEN inhibitor, an AKT inhibitor, or an mTOR inhibitor(e.g., mTORC1 inhibitor or mTORC2 inhibitor). In some embodiments, theanti-cancer agent is JAB-3312.

In some embodiments, an anti-cancer agent is a SOS1 inhibitor. In someembodiments, the SOS1 inhibitor is selected from those disclosed in WO2021173524, WO 2021130731, WO 2021127429, WO 2021092115, WO 2021105960,WO 2021074227, WO 2020180768, WO 2020180770, WO 2020173935, WO2020146470, WO 2019201848, WO 2019122129, WO 2018172250, and WO2018115380, or a pharmaceutically acceptable salt, solvate, isomer(e.g., stereoisomer), prodrug, or tautomer thereof.

In some embodiments, an anti-cancer agent is an additional Ras inhibitoror a Ras vaccine, or another therapeutic modality designed to directlyor indirectly decrease the oncogenic activity of Ras. In someembodiments, an anti-cancer agent is an additional Ras inhibitor. Insome embodiments, the Ras inhibitor targets Ras in its active, orGTP-bound state. In some embodiments, the Ras inhibitor targets Ras inits inactive, or GDP-bound state. In some embodiments, the Ras inhibitoris, such as an inhibitor of K-Ras G12C, such as AMG 510 (sotorasib),MRTX1257, MRTX849 (adagrasib), JNJ-74699157, LY3499446, ARS-1620,ARS-853, BPI-421286, LY3537982, JDQ443, JAB-21000, RMC-6291 or GDC-6036,or a pharmaceutically acceptable salt, solvate, isomer (e.g.,stereoisomer), prodrug, or tautomer thereof. In some embodiments, theRas inhibitor is an inhibitor of K-Ras G12D, such as MRTX1133 orJAB-22000, or a pharmaceutically acceptable salt, solvate, isomer (e.g.,stereoisomer), prodrug, or tautomer thereof. In some embodiments, theRas inhibitor is a K-Ras G12V inhibitor, such as JAB-23000, or apharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer),prodrug, or tautomer thereof. In some embodiments, the Ras inhibitor isRMC-6236, or a pharmaceutically acceptable salt, solvate, isomer (e.g.,stereoisomer), prodrug, or tautomer thereof. In some embodiments, theRas inhibitor is selected from a Ras(ON) inhibitor disclosed in thefollowing, incorporated herein by reference in their entireties, or apharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer),prodrug, or tautomer thereof: WO 2021091982, WO 2021091967, WO2021091956 and WO 2020132597. Other examples of Ras inhibitors that maybe combined with a Ras inhibitor of the present invention are providedin the following, incorporated herein by reference in their entireties:WO 2021173923, WO 2021169990, WO 2021169963, WO 2021168193, WO2021158071, WO 2021155716, WO 2021152149, WO 2021150613, WO 2021147967,WO 2021147965, WO 2021143693, WO 2021142252, WO 2021141628, WO2021139748, WO 2021139678, WO 2021129824, WO 2021129820, WO 2021127404,WO 2021126816, WO 2021126799, WO 2021124222, WO 2021121371, WO2021121367, WO 2021121330, WO 2020050890, WO 2020047192, WO 2020035031,WO 2020028706, WO 2019241157, WO 2019232419, WO 2019217691, WO2019217307, WO 2019215203, WO 2019213526, WO 2019213516, WO 2019155399,WO 2019150305, WO 2019110751, WO 2019099524, WO 2019051291, WO2018218070, WO 2018217651, WO 2018218071, WO 2018218069, WO 2018206539,WO 2018143315, WO 2018140600, WO 2018140599, WO 2018140598, WO2018140514, WO 2018140513, WO 2018140512, WO 2018119183, WO 2018112420,WO 2018068017, WO 2018064510, WO 2017201161, WO 2017172979, WO2017100546, WO 2017087528, WO 2017058807, WO 2017058805, WO 2017058728,WO 2017058902, WO 2017058792, WO 2017058768, WO 2017058915, WO2017015562, WO 2016168540, WO 2016164675, WO 2016049568, WO 2016049524,WO 2015054572, WO 2014152588, WO 2014143659, and WO 2013155223, or apharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer),prodrug, or tautomer thereof.

In some embodiments, a therapeutic agent that may be combined with acompound of the present invention is an inhibitor of the MAP kinase(MAPK) pathway (or “MAPK inhibitor”). MAPK inhibitors include, but arenot limited to, one or more MAPK inhibitor described in Cancers (Basel)2015 September; 7(3): 1758-1784. For example, the MAPK inhibitor may beselected from one or more of trametinib, binimetinib, selumetinib,cobimetinib, LErafAON (NeoPharm), ISIS 5132; vemurafenib, pimasertib,TAK733, R04987655 (CH4987655); CI-1040; PD-0325901; CH5126766; MAP855;AZD6244; refametinib (RDEA 119/BAY 86-9766); GDC-0973/XL581; AZD8330(ARRY-424704/ARRY-704); R05126766 (Roche, described in PLoS One. 2014Nov. 25; 9(11)); and GSK1120212 (or JTP-74057, described in Clin CancerRes. 2011 Mar. 1; 17(5):989-1000). The MAPK inhibitor may be PLX8394,LXH254, GDC-5573, or LY3009120.

In some embodiments, an anti-cancer agent is a disrupter or inhibitor ofthe RAS-RAF-ERK or PI3K-AKT-TOR or PI3K-AKT signaling pathways. ThePI3K/AKT inhibitor may include, but is not limited to, one or morePI3K/AKT inhibitor described in Cancers (Basel) 2015 September; 7(3):1758-1784. For example, the PI3K/AKT inhibitor may be selected from oneor more of NVP-BEZ235; BGT226; XL765/SAR245409; SF1126; GDC-0980;PI-103; PF-04691502; PKI-587; GSK2126458.

In some embodiments, an anti-cancer agent is a PD-1 or PD-L1 antagonist.

In some embodiments, additional therapeutic agents include ALKinhibitors, HER2 inhibitors, EGFR inhibitors, IGF-1R inhibitors, MEKinhibitors, PI3K inhibitors, AKT inhibitors, TOR inhibitors, MCL-1inhibitors, BCL-2 inhibitors, SHP2 inhibitors, proteasome inhibitors,and immune therapies. In some embodiments, a therapeutic agent may be apan-RTK inhibitor, such as afatinib.

IGF-1R inhibitors include linsitinib, or a pharmaceutically acceptablesalt thereof.

EGFR inhibitors include, but are not limited to, small moleculeantagonists, antibody inhibitors, or specific antisense nucleotide orsiRNA. Useful antibody inhibitors of EGFR include cetuximab (Erbitux®),panitumumab (Vectibix®), zalutumumab, nimotuzumab, and matuzumab.Further antibody-based EGFR inhibitors include any anti-EGFR antibody orantibody fragment that can partially or completely block EGFR activationby its natural ligand. Non-limiting examples of antibody-based EGFRinhibitors include those described in Modjtahedi et al., Br. J. Cancer1993, 67:247-253; Teramoto et al., Cancer 1996, 77:639-645; Goldstein etal., Clin. Cancer Res. 1995, 1:1311-1318; Huang et al., 1999, CancerRes. 15:59(8):1935-40; and Yang et al., Cancer Res. 1999, 59:1236-1243.The EGFR inhibitor can be monoclonal antibody Mab E7.6.3 (Yang, 1999supra), or Mab C₂₂₅ (ATCC Accession No. HB-8508), or an antibody orantibody fragment having the binding specificity thereof.

Small molecule antagonists of EGFR include gefitinib (Iressa®),erlotinib (Tarceva®), and lapatinib (TykerB®). See, e.g., Yan et al.,Pharmacogenetics and Pharmacogenomics In Oncology Therapeutic AntibodyDevelopment, BioTechniques 2005, 39(4):565-8; and Paez et al., EGFRMutations In Lung Cancer Correlation With Clinical Response To GefitinibTherapy, Science 2004, 304(5676):1497-500. In some embodiments, the EGFRinhibitor is osimertinib (Tagrisso®). Further non-limiting examples ofsmall molecule EGFR inhibitors include any of the EGFR inhibitorsdescribed in the following patent publications, and all pharmaceuticallyacceptable salts of such EGFR inhibitors: EP 0520722; EP 0566226;WO96/33980; U.S. Pat. No. 5,747,498; WO96/30347; EP 0787772; WO97/30034;WO97/30044; WO97/38994; WO97/49688; EP 837063; WO98/02434; WO97/38983;WO95/19774; WO95/19970; WO97/13771; WO98/02437; WO98/02438; WO97/32881;DE 19629652; WO98/33798; WO97/32880; WO97/32880; EP 682027; WO97/02266;WO97/27199; WO98/07726; WO97/34895; WO96/31510; WO98/14449; WO98/14450;WO98/14451; WO95/09847; WO97/19065; WO98/17662; U.S. Pat. Nos.5,789,427; 5,650,415; 5,656,643; WO99/35146; WO99/35132; WO99/07701; andWO92/20642. Additional non-limiting examples of small molecule EGFRinhibitors include any of the EGFR inhibitors described in Traxler etal., Exp. Opin. Ther. Patents 1998, 8(12):1599-1625. In someembodiments, an EGFR inhibitor is an ERBB inhibitor. In humans, the ERBBfamily contains HER1 (EGFR, ERBB1), HER2 (NEU, ERBB2), HER3 (ERBB3), andHER (ERBB4).

MEK inhibitors include, but are not limited to, pimasertib, selumetinib,cobimetinib (Cotellic®), trametinib (Mekinist®), and binimetinib(Mektovi®). In some embodiments, a MEK inhibitor targets a MEK mutationthat is a Class I MEK1 mutation selected from D67N; P124L; P124S; andL177V. In some embodiments, the MEK mutation is a Class II MEK1 mutationselected from ΔE51-Q58; ΔF53-Q58; E203K; L177M; C121S; F53L; K57E; Q56P;and K57N.

PI3K inhibitors include, but are not limited to, wortmannin;17-hydroxywortmannin analogs described in WO06/044453;4-[2-(1H-Indazol-4-yl)-6-[[4-(methylsulfonyl)piperazin-1-yl]methyl]thieno[3,2-d]pyrimidin-4-yl]morpholine(also known as pictilisib or GDC-0941 and described in WO09/036082 andWO09/055730);2-methyl-2-[4-[3-methyl-2-oxo-8-(quinolin-3-yl)-2,3-dihydroimidazo[4,5-c]quinolin-1-yl]phenyl]propionitrile(also known as BEZ 235 or NVP-BEZ 235, and described in WO06/122806);(S)—I-(4-((2-(2-aminopyrimidin-5-yl)-7-methyl-4-morpholinothieno[3,2-d]pyrimidin-6-yl)methyl)piperazin-1-yl)-2-hydroxypropan-1-one(described in WO08/070740); LY294002(2-(4-morpholinyl)-8-phenyl-4H-I-benzopyran-4-one (available from AxonMedchem); PI 103 hydrochloride(3-[4-(4-morpholinylpyrido-[3′,2′:4,5]furo[3,2-d]pyrimidin-2-yl] phenolhydrochloride (available from Axon Medchem); PIK 75(2-methyl-5-nitro-2-[(6-bromoimidazo[1,2-a]pyridin-3-yl)methylene]-1-methylhydrazide-benzenesulfonicacid, monohydrochloride) (available from Axon Medchem); PIK 90(N-(7,8-dimethoxy-2,3-dihydro-imidazo[I,2-c]quinazolin-5-yl)-nicotinamide(available from Axon Medchem); AS-252424(5-[I-[5-(4-fluoro-2-hydroxy-phenyl)-furan-2-yl]-meth-(Z)-ylidene]-thiazolidine-2,4-dione(available from Axon Medchem); TGX-221(7-methyl-2-(4-morpholinyl)-9-[1-(phenylamino)ethyl]-4H-pyrido-[1,2-a]pyrirnidin-4-one(available from Axon Medchem); XL-765; and XL-147. Other PI3K inhibitorsinclude demethoxyviridin, perifosine, CAL101, PX-866, BEZ235, SF1126,INK1117, IPI-145, BKM120, XL147, XL765, Palomid 529, GSK1059615,ZSTK474, PWT33597, IC87114, TGI 00-115, CAL263, PI-103, GNE-477,CUDC-907, and AEZS-136.

AKT inhibitors include, but are not limited to, Akt-1-1 (inhibits Aktl)(Barnett et at, Biochem. J. 2005, 385 (Pt. 2): 399-408); Akt-1-1,2(inhibits Akl and 2) (Barnett et at, Biochem. J. 2005, 385 (Pt. 2):399-408); API-59CJ-Ome (e.g., Jin et al., Br. J. Cancer 2004,91:1808-12); 1-H-imidazo[4,5-c]pyridinyl compounds (e.g., WO 05/011700);indole-3-carbinol and derivatives thereof (e.g., U.S. Pat. No.6,656,963; Sarkar and Li J Nutr. 2004, 134(12 Suppl):3493S-3498S);perifosine (e.g., interferes with Akt membrane localization;Dasmahapatra et al. Clin. Cancer Res. 2004, 10(15):5242-52);phosphatidylinositol ether lipid analogues (e.g., Gills and DennisExpert. Opin. Investig. Drugs 2004, 13:787-97); and triciribine (TCN orAPI-2 or NCI identifier: NSC 154020; Yang et at, Cancer Res. 2004,64:4394-9).

mTOR inhibitors include, but are not limited to, ATP-competitivemTORC1/mTORC2 inhibitors, e.g., PI-103, PP242, PP30; Torin 1; FKBP12enhancers; 4H-1-benzopyran-4-one derivatives; and rapamycin (also knownas sirolimus) and derivatives thereof, including: temsirolimus(Torisel®); everolimus (Afinitor®; WO94/09010); ridaforolimus (alsoknown as deforolimus or AP23573); rapalogs, e.g., as disclosed inWO98/02441 and WO01/14387, e.g. AP23464 and AP23841;40-(2-hydroxyethyl)rapamycin;40-[3-hydroxy(hydroxymethyl)methylpropanoate]-rapamycin (also known asCC1779); 40-epi-(tetrazolyt)-rapamycin (also called ABT578);32-deoxorapamycin; 16-pentynyloxy-32(S)-dihydrorapanycin; derivativesdisclosed in WO05/005434; derivatives disclosed in U.S. Pat. Nos.5,258,389, 5,118,677, 5,118,678, 5,100,883, 5,151,413, 5,120,842, and5,256,790, and in WO94/090101, WO92/05179, WO93/111130, WO94/02136,WO94/02485, WO95/14023, WO94/02136, WO95/16691, WO96/41807, WO96/41807,and WO2018204416; and phosphorus-containing rapamycin derivatives (e.g.,WO05/016252). In some embodiments, the mTOR inhibitor is a bistericinhibitor (see, e.g., WO2018204416, WO2019212990 and WO2019212991), suchas RMC-5552, having the structure

BRAF inhibitors that may be used in combination with compounds of theinvention include, for example, vemurafenib, dabrafenib, andencorafenib. A BRAF may comprise a Class 3 BRAF mutation. In someembodiments, the Class 3 BRAF mutation is selected from one or more ofthe following amino acid substitutions in human BRAF: D287H; P367R;V459L; G466V; G466E; G466A; S467L; G469E; N581S; N5811; D594N; D594G;D594A; D594H; F595L; G596D; G596R and A762E.

MCL-1 inhibitors include, but are not limited to, AMG-176, MIK665, andS63845. The myeloid cell leukemia-1 (MCL-1) protein is one of the keyanti-apoptotic members of the B-cell lymphoma-2 (BCL-2) protein family.Over-expression of MCL-1 has been closely related to tumor progressionas well as to resistance, not only to traditional chemotherapies butalso to targeted therapeutics including BCL-2 inhibitors such asABT-263.

In some embodiments, the additional therapeutic agent is a SHP2inhibitor. SHP2 is a non-receptor protein tyrosine phosphatase encodedby the PTPN11 gene that contributes to multiple cellular functionsincluding proliferation, differentiation, cell cycle maintenance andmigration. SHP2 has two N-terminal Src homology 2 domains (N—SH2 andC—SH2), a catalytic domain (PTP), and a C-terminal tail. The two SH2domains control the subcellular localization and functional regulationof SHP2. The molecule exists in an inactive, self-inhibited conformationstabilized by a binding network involving residues from both the N—SH2and PTP domains. Stimulation by, for example, cytokines or growthfactors acting through receptor tyrosine kinases (RTKs) leads toexposure of the catalytic site resulting in enzymatic activation ofSHP2.

SHP2 is involved in signaling through the RAS-mitogen-activated proteinkinase (MAPK), the JAK-STAT or the phosphoinositol 3-kinase-AKTpathways. Mutations in the PTPN11 gene and subsequently in SHP2 havebeen identified in several human developmental diseases, such as NoonanSyndrome and Leopard Syndrome, as well as human cancers, such asjuvenile myelomonocytic leukemia, neuroblastoma, melanoma, acute myeloidleukemia and cancers of the breast, lung and colon. Some of thesemutations destabilize the auto-inhibited conformation of SHP2 andpromote autoactivation or enhanced growth factor driven activation ofSHP2. SHP2, therefore, represents a highly attractive target for thedevelopment of novel therapies for the treatment of various diseasesincluding cancer. A SHP2 inhibitor (e.g., RMC-4550 or SHP099) incombination with a RAS pathway inhibitor (e.g., a MEK inhibitor) havebeen shown to inhibit the proliferation of multiple cancer cell lines invitro (e.g., pancreas, lung, ovarian and breast cancer). Thus,combination therapy involving a SHP2 inhibitor with a RAS pathwayinhibitor could be a general strategy for preventing tumor resistance ina wide range of malignancies.

Non-limiting examples of such SHP2 inhibitors that are known in the art,include: Chen et al. Mol Pharmacol. 2006, 70, 562; Sarver et al., J.Med. Chem. 2017, 62, 1793; Xie et al., J. Med. Chem. 2017, 60, 113734;and Igbe et al., Oncotarget, 2017, 8, 113734; and PCT applications: WO2021149817, WO 2021148010, WO 2021147879, WO 2021143823, WO 2021143701,WO 2021143680, WO 2021121397, WO 2021119525, WO 2021115286, WO2021110796, WO 2021088945, WO 2021073439, WO 2021061706, WO 2021061515,WO 2021043077, WO 2021033153, WO 2021028362, WO 2021033153, WO2021028362, WO 2021018287, WO 2020259679, WO 2020249079, WO 2020210384,WO 2020201991, WO 2020181283, WO 2020177653, WO 2020165734, WO2020165733, WO 2020165732, WO 2020156243, WO 2020156242, WO 2020108590,WO 2020104635, WO 2020094104, WO 2020094018, WO 2020081848, WO2020073949, WO 2020073945, WO 2020072656, WO 2020065453, WO 2020065452,WO 2020063760, WO 2020061103, WO 2020061101, WO 2020033828, WO2020033286, WO 2020022323, WO 2019233810, WO 2019213318, WO 2019183367,WO 2019183364, WO 2019182960, WO 2019167000, WO 2019165073, WO2019158019, WO 2019152454, WO 2019051469, WO 2019051084, WO 2018218133,WO 2018172984, WO 2018160731, WO 2018136265, WO 2018136264, WO2018130928, WO 2018129402, WO 2018081091, WO 2018057884, WO 2018013597,WO 2017216706, WO 2017211303, WO 2017210134, WO 2017156397, WO2017100279, WO 2017079723, WO 2017078499, WO 2016203406, WO 2016203405,WO 2016203404, WO 2016196591, WO 2016191328, WO 2015107495, WO2015107494, WO 2015107493, WO 2014176488, WO 2014113584, US 20210085677,U.S. Ser. No. 10/858,359, U.S. Ser. No. 10/934,302, U.S. Ser. No.10/954,243, U.S. Ser. No. 10/988,466, U.S. Ser. No. 11/001,561, U.S.Ser. No. 11/033,547, U.S. Ser. No. 11/034,705, or U.S. Ser. No.11/044,675, or a pharmaceutically acceptable salt, solvate, isomer(e.g., stereoisomer), prodrug, or tautomer thereof, each of which isincorporated herein by reference.

In some embodiments, a SHP2 inhibitor binds in the active site. In someembodiments, a SHP2 inhibitor is a mixed-type irreversible inhibitor. Insome embodiments, a SHP2 inhibitor binds an allosteric site e.g., anon-covalent allosteric inhibitor. In some embodiments, a SHP2 inhibitoris a covalent SHP2 inhibitor, such as an inhibitor that targets thecysteine residue (C333) that lies outside the phosphatase's active site.In some embodiments a SHP2 inhibitor is a reversible inhibitor. In someembodiments, a SHP2 inhibitor is an irreversible inhibitor. In someembodiments, the SHP2 inhibitor is SHP099. In some embodiments, the SHP2inhibitor is TNO155, having the structure

or a pharmaceutically acceptable salt, solvate, isomer (e.g.,stereoisomer), prodrug, or tautomer thereof.In some embodiments, the SHP2 inhibitor is RMC-4550, having thestructure

or a pharmaceutically acceptable salt, solvate, isomer (e.g.,stereoisomer), prodrug, or tautomer thereof. In some embodiments, theSHP2 inhibitor is RMC-4630, having the structure:

or a pharmaceutically acceptable salt, solvate, isomer (e.g.,stereoisomer), prodrug, or tautomer thereof. In some embodiments, theSHP2 inhibitor is JAB-3068, having the structure

or a pharmaceutically acceptable salt, solvate, isomer (e.g.,stereoisomer), prodrug, or tautomer thereof. In some embodiments, theSHP2 inhibitor is JAB-3312. In some embodiments, the SHP2 inhibitor isthe following compound,

or a pharmaceutically acceptable salt, solvate, isomer (e.g.,stereoisomer), prodrug, or tautomer thereof. In some embodiments, theSHP2 inhibitor is RLY-1971, having the structure

or a pharmaceutically acceptable salt, solvate, isomer (e.g.,stereoisomer), prodrug, or tautomer thereof. In some embodiments, theSHP2 inhibitor is ERAS-601, or a pharmaceutically acceptable salt,solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof. Insome embodiments, the SHP2 inhibitor is BBP-398, or a pharmaceuticallyacceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, ortautomer thereof. In some embodiments, the SHP2 inhibitor is SH3809. Insome embodiments, the SHP2 inhibitor is PF-07284892, or apharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer),prodrug, or tautomer thereof.

In some embodiments, the additional therapeutic agent is selected fromthe group consisting of a MEK inhibitor, HER2 inhibitor, a SHP2inhibitor, CDK4/6 inhibitor, an mTOR inhibitor, a SOS1 inhibitor, and aPD-L1 inhibitor. In some embodiments, the additional therapeutic agentis selected from the group consisting of a MEK inhibitor, a SHP2inhibitor, and a PD-L1 inhibitor. See, e.g., Hallin et al., CancerDiscovery, DOI: 10.1158/2159-8290 (Oct. 28, 2019) and Canon et al.,Nature, 575:217 (2019). In some embodiments, a Ras inhibitor of thepresent invention is used in combination with a MEK inhibitor and a SOS1inhibitor. In some embodiments, a Ras inhibitor of the present inventionis used in combination with a PD-L1 inhibitor and a SOS1 inhibitor. Insome embodiments, a Ras inhibitor of the present invention is used incombination with a PD-L1 inhibitor and a SHP2 inhibitor. In someembodiments, a Ras inhibitor of the present invention is used incombination with a MEK inhibitor and a SHP2 inhibitor. In someembodiments, the cancer is colorectal cancer and the treatment comprisesadministration of a Ras inhibitor of the present invention incombination with a second or third therapeutic agent.

Proteasome inhibitors include, but are not limited to, carfilzomib(Kyprolis®), bortezomib (Velcade®), and oprozomib.

Immune therapies include, but are not limited to, monoclonal antibodies,immunomodulatory imides (IMiDs), GITR agonists, genetically engineeredT-cells (e.g., CAR-T cells), bispecific antibodies (e.g., BiTEs), andanti-PD-1, anti-PD-L1, anti-CTLA4, anti-LAGI, and anti-OX40 agents).

Immunomodulatory agents (IMiDs) are a class of immunomodulatory drugs(drugs that adjust immune responses) containing an imide group. The IMiDclass includes thalidomide and its analogues (lenalidomide,pomalidomide, and apremilast).

Exemplary anti-PD-1 antibodies and methods for their use are describedby Goldberg et al., Blood 2007, 110(1):186-192; Thompson et al., Clin.Cancer Res. 2007, 13(6):1757-1761; and WO06/121168 A1), as well asdescribed elsewhere herein.

GITR agonists include, but are not limited to, GITR fusion proteins andanti-GITR antibodies (e.g., bivalent anti-GITR antibodies), such as, aGITR fusion protein described in U.S. Pat. Nos. 6,111,090, 8,586,023,WO2010/003118 and WO2011/090754; or an anti-GITR antibody described,e.g., in U.S. Pat. No. 7,025,962, EP 1947183, U.S. Pat. Nos. 7,812,135,8,388,967, 8,591,886, 7,618,632, EP 1866339, and WO2011/028683,WO2013/039954, WO05/007190, WO07/133822, WO05/055808, WO99/40196,WO01/03720, WO99/20758, WO06/083289, WO05/115451, and WO2011/051726.

Another example of a therapeutic agent that may be used in combinationwith the compounds of the invention is an anti-angiogenic agent.Anti-angiogenic agents are inclusive of, but not limited to, in vitrosynthetically prepared chemical compositions, antibodies, antigenbinding regions, radionuclides, and combinations and conjugates thereof.An anti-angiogenic agent can be an agonist, antagonist, allostericmodulator, toxin or, more generally, may act to inhibit or stimulate itstarget (e.g., receptor or enzyme activation or inhibition), and therebypromote cell death or arrest cell growth. In some embodiments, the oneor more additional therapies include an anti-angiogenic agent.

Anti-angiogenic agents can be MMP-2 (matrix-metalloproteinase 2)inhibitors, MMP-9 (matrix-metalloprotienase 9) inhibitors, and COX-II(cyclooxygenase 11) inhibitors. Non-limiting examples of anti-angiogenicagents include rapamycin, temsirolimus (CCI-779), everolimus (RAD001),sorafenib, sunitinib, and bevacizumab. Examples of useful COX-IIinhibitors include alecoxib, valdecoxib, and rofecoxib. Examples ofuseful matrix metalloproteinase inhibitors are described in WO96/33172,WO96/27583, WO98/07697, WO98/03516, WO98/34918, WO98/34915, WO98/33768,WO98/30566, WO90/05719, WO99/52910, WO99/52889, WO99/29667, WO99007675,EP0606046, EP0780386, EP1786785, EP1181017, EP0818442, EP1004578, andUS20090012085, and U.S. Pat. Nos. 5,863,949 and 5,861,510. PreferredMMP-2 and MMP-9 inhibitors are those that have little or no activityinhibiting MMP-1. More preferred, are those that selectively inhibitMMP-2 or AMP-9 relative to the other matrix-metalloproteinases (i.e.,MAP-1, MMP-3, MMP-4, MMP-5, MMP-6, MMP-7, MMP-8, MMP-10, MMP-11, MMP-12,and MMP-13). Some specific examples of MMP inhibitors are AG-3340, RO32-3555, and RS 13-0830.

Further exemplary anti-angiogenic agents include KDR (kinase domainreceptor) inhibitory agents (e.g., antibodies and antigen bindingregions that specifically bind to the kinase domain receptor), anti-VEGFagents (e.g., antibodies or antigen binding regions that specificallybind VEGF (e.g., bevacizumab), or soluble VEGF receptors or a ligandbinding region thereof) such as VEGF-TRAP™, and anti-VEGF receptoragents (e.g., antibodies or antigen binding regions that specificallybind thereto), EGFR inhibitory agents (e.g., antibodies or antigenbinding regions that specifically bind thereto) such as Vectibix®(panitumumab), erlotinib (Tarceva®), anti-AngI and anti-Ang2 agents(e.g., antibodies or antigen binding regions specifically bindingthereto or to their receptors, e.g., Tie2/Tek), and anti-Tie2 kinaseinhibitory agents (e.g., antibodies or antigen binding regions thatspecifically bind thereto). Other anti-angiogenic agents includeCampath, IL-8, B-FGF, Tek antagonists (US2003/0162712; U.S. Pat. No.6,413,932), anti-TWEAK agents (e.g., specifically binding antibodies orantigen binding regions, or soluble TWEAK receptor antagonists; see U.S.Pat. No. 6,727,225), ADAM distintegrin domain to antagonize the bindingof integrin to its ligands (US 2002/0042368), specifically bindinganti-eph receptor or anti-ephrin antibodies or antigen binding regions(U.S. Pat. Nos. 5,981,245; 5,728,813; 5,969,110; 6,596,852; 6,232,447;6,057,124 and patent family members thereof), and anti-PDGF-BBantagonists (e.g., specifically binding antibodies or antigen bindingregions) as well as antibodies or antigen binding regions specificallybinding to PDGF-BB ligands, and PDGFR kinase inhibitory agents (e.g.,antibodies or antigen binding regions that specifically bind thereto).Additional anti-angiogenic agents include: SD-7784 (Pfizer, USA);cilengitide (Merck KGaA, Germany, EPO 0770622); pegaptanib octasodium,(Gilead Sciences, USA); Alphastatin, (BioActa, UK); M-PGA, (Celgene,USA, U.S. Pat. No. 5,712,291); ilomastat, (Arriva, USA, U.S. Pat. No.5,892,112); emaxanib, (Pfizer, USA, U.S. Pat. No. 5,792,783); vatalanib,(Novartis, Switzerland); 2-methoxyestradiol (EntreMed, USA); TLC ELL-12(Elan, Ireland); anecortave acetate (Alcon, USA); alpha-D148 Mab (Amgen,USA); CEP-7055 (Cephalon, USA); anti-Vn Mab (Crucell, Netherlands),DACantiangiogenic (ConjuChem, Canada); Angiocidin (InKinePharmaceutical, USA); KM-2550 (Kyowa Hakko, Japan); SU-0879 (Pfizer,USA); CGP-79787 (Novartis, Switzerland, EP 0970070); ARGENT technology(Ariad, USA); YIGSR-Stealth (Johnson & Johnson, USA); fibrinogen-Efragment (BioActa, UK); angiogenic inhibitor (Trigen, UK); TBC-1635(Encysive Pharmaceuticals, USA); SC-236 (Pfizer, USA); ABT-567 (Abbott,USA); Metastatin (EntreMed, USA); maspin (Sosei, Japan);2-methoxyestradiol (Oncology Sciences Corporation, USA); ER-68203-00 (IVAX, USA); BeneFin (Lane Labs, USA); Tz-93 (Tsumura, Japan); TAN-1120(Takeda, Japan); FR-111142 (Fujisawa, Japan, JP 02233610); plateletfactor 4 (RepliGen, USA, EP 407122); vascular endothelial growth factorantagonist (Borean, Denmark); bevacizumab (pINN) (Genentech, USA);angiogenic inhibitors (SUGEN, USA); XL 784 (Exelixis, USA); XL 647(Exelixis, USA); MAb, alpha5beta3 integrin, second generation (AppliedMolecular Evolution, USA and Medlmmune, USA); enzastaurin hydrochloride(Lilly, USA); CEP 7055 (Cephalon, USA and Sanofi-Synthelabo, France); BC1 (Genoa Institute of Cancer Research, Italy); rBPI 21 and BPI-derivedantiangiogenic (XOMA, USA); PI 88 (Progen, Australia); cilengitide(Merck KGaA, German; Munich Technical University, Germany, ScrippsClinic and Research Foundation, USA); AVE 8062 (Ajinomoto, Japan); AS1404 (Cancer Research Laboratory, New Zealand); SG 292, (Telios, USA);Endostatin (Boston Childrens Hospital, USA); ATN 161 (Attenuon, USA);2-methoxyestradiol (Boston Childrens Hospital, USA); ZD 6474,(AstraZeneca, UK); ZD 6126, (Angiogene Pharmaceuticals, UK); PPI 2458,(Praecis, USA); AZD 9935, (AstraZeneca, UK); AZD 2171, (AstraZeneca,UK); vatalanib (pINN), (Novartis, Switzerland and Schering AG, Germany);tissue factor pathway inhibitors, (EntreMed, USA); pegaptanib (Pinn),(Gilead Sciences, USA); xanthorrhizol, (Yonsei University, South Korea);vaccine, gene-based, VEGF-2, (Scripps Clinic and Research Foundation,USA); SPV5.2, (Supratek, Canada); SDX 103, (University of California atSan Diego, USA); PX478, (ProIX, USA); METASTATIN, (EntreMed, USA);troponin I, (Harvard University, USA); SU 6668, (SUGEN, USA); OXI 4503,(OXiGENE, USA); o-guanidines, (Dimensional Pharmaceuticals, USA);motuporamine C, (British Columbia University, Canada); CDP 791,(Celltech Group, UK); atiprimod (pINN), (GlaxoSmithKline, UK); E 7820,(Eisai, Japan); CYC 381, (Harvard University, USA); AE 941, (Aeterna,Canada); vaccine, angiogenic, (EntreMed, USA); urokinase plasminogenactivator inhibitor, (Dendreon, USA); oglufanide (pINN), (Melmotte,USA); HIF-lalfa inhibitors, (Xenova, UK); CEP 5214, (Cephalon, USA); BAYRES 2622, (Bayer, Germany); Angiocidin, (InKine, USA); A6, (Angstrom,USA); KR 31372, (Korea Research Institute of Chemical Technology, SouthKorea); GW 2286, (GlaxoSmithKline, UK); EHT 0101, (ExonHit, France); CP868596, (Pfizer, USA); CP 564959, (OSI, USA); CP 547632, (Pfizer, USA);786034, (GlaxoSmithKline, UK); KRN 633, (Kirin Brewery, Japan); drugdelivery system, intraocular, 2-methoxyestradiol; anginex (MaastrichtUniversity, Netherlands, and Minnesota University, USA); ABT 510(Abbott, USA); AAL 993 (Novartis, Switzerland); VEGI (ProteomTech, USA);tumor necrosis factor-alpha inhibitors; SU 11248 (Pfizer, USA and SUGENUSA); ABT 518, (Abbott, USA); YH16 (Yantai Rongchang, China); S-3APG(Boston Childrens Hospital, USA and EntreMed, USA); MAb, KDR (ImCloneSystems, USA); MAb, alpha5 beta (Protein Design, USA); KDR kinaseinhibitor (Celltech Group, UK, and Johnson & Johnson, USA); GFB 116(South Florida University, USA and Yale University, USA); CS 706(Sankyo, Japan); combretastatin A4 prodrug (Arizona State University,USA); chondroitinase AC (IBEX, Canada); BAY RES 2690 (Bayer, Germany);AGM 1470 (Harvard University, USA, Takeda, Japan, and TAP, USA); AG13925 (Agouron, USA); Tetrathiomolybdate (University of Michigan, USA);GCS 100 (Wayne State University, USA) CV 247 (Ivy Medical, UK); CKD 732(Chong Kun Dang, South Korea); irsogladine, (Nippon Shinyaku, Japan); RG13577 (Aventis, France); WX 360 (Wilex, Germany); squalamine, (Genaera,USA); RPI 4610 (Sirna, USA); heparanase inhibitors (InSight, Israel); KL3106 (Kolon, South Korea); Honokiol (Emory University, USA); ZK CDK(Schering AG, Germany); ZK Angio (Schering AG, Germany); ZK 229561(Novartis, Switzerland, and Schering AG, Germany); XMP 300 (XOMA, USA);VGA 1102 (Taisho, Japan); VE-cadherin-2 antagonists (ImClone Systems,USA); Vasostatin (National Institutes of Health, USA); Flk-1 (ImCloneSystems, USA); TZ 93 (Tsumura, Japan); TumStatin (Beth Israel Hospital,USA); truncated soluble FLT 1 (vascular endothelial growth factorreceptor 1) (Merck & Co, USA); Tie-2 ligands (Regeneron, USA); andthrombospondin 1 inhibitor (Allegheny Health, Education and ResearchFoundation, USA).

Further examples of therapeutic agents that may be used in combinationwith compounds of the invention include agents (e.g., antibodies,antigen binding regions, or soluble receptors) that specifically bindand inhibit the activity of growth factors, such as antagonists ofhepatocyte growth factor (HGF, also known as Scatter Factor), andantibodies or antigen binding regions that specifically bind itsreceptor, c-Met.

Another example of a therapeutic agent that may be used in combinationwith compounds of the invention is an autophagy inhibitor. Autophagyinhibitors include, but are not limited to chloroquine, 3-methyladenine,hydroxychloroquine (Plaquenil™), bafilomycin A1,5-amino-4-imidazolecarboxamide riboside (AICAR), okadaic acid, autophagy-suppressive algaltoxins which inhibit protein phosphatases of type 2A or type 1,analogues of cAMP, and drugs which elevate cAMP levels such asadenosine, LY204002, N6-mercaptopurine riboside, and vinblastine. Inaddition, antisense orsiRNAthat inhibits expression of proteinsincluding but not limited to ATG5 (which are implicated in autophagy),may also be used. In some embodiments, the one or more additionaltherapies include an autophagy inhibitor.

Another example of a therapeutic agent that may be used in combinationwith compounds of the invention is an anti-neoplastic agent. In someembodiments, the one or more additional therapies include ananti-neoplastic agent. Non-limiting examples of anti-neoplastic agentsinclude acemannan, aclarubicin, aldesleukin, alemtuzumab, alitretinoin,altretamine, amifostine, aminolevulinic acid, amrubicin, amsacrine,anagrelide, anastrozole, ancer, ancestim, arglabin, arsenic trioxide,BAM-002 (Novelos), bexarotene, bicalutamide, broxuridine, capecitabine,celmoleukin, cetrorelix, cladribine, clotrimazole, cytarabine ocfosfate,DA 3030 (Dong-A), daclizumab, denileukin diftitox, deslorelin,dexrazoxane, dilazep, docetaxel, docosanol, doxercalciferol,doxifluridine, doxorubicin, bromocriptine, carmustine, cytarabine,fluorouracil, HIT diclofenac, interferon alfa, daunorubicin,doxorubicin, tretinoin, edelfosine, edrecolomab, eflornithine, emitefur,epirubicin, epoetin beta, etoposide phosphate, exemestane, exisulind,fadrozole, filgrastim, finasteride, fludarabine phosphate, formestane,fotemustine, gallium nitrate, gemcitabine, gemtuzumab zogamicin,gimeracil/oteracil/tegafur combination, glycopine, goserelin,heptaplatin, human chorionic gonadotropin, human fetal alphafetoprotein, ibandronic acid, idarubicin, (imiquimod, interferon alfa,interferon alfa, natural, interferon alfa-2, interferon alfa-2a,interferon alfa-2b, interferon alfa-NI, interferon alfa-n3, interferonalfacon-1, interferon alpha, natural, interferon beta, interferonbeta-Ia, interferon beta-Ib, interferon gamma, natural interferongamma-Ia, interferon gamma-Ib, interleukin-1 beta, iobenguane,irinotecan, irsogladine, lanreotide, LC 9018 (Yakult), leflunomide,lenograstim, lentinan sulfate, letrozole, leukocyte alpha interferon,leuprorelin, levamisole+fluorouracil, liarozole, lobaplatin, lonidamine,lovastatin, masoprocol, melarsoprol, metoclopramide, mifepristone,miltefosine, mirimostim, mismatched double stranded RNA, mitoguazone,mitolactol, mitoxantrone, molgramostim, nafarelin, naloxone+pentazocine,nartograstim, nedaplatin, nilutamide, noscapine, novel erythropoiesisstimulating protein, NSC 631570 octreotide, oprelvekin, osaterone,oxaliplatin, paclitaxel, pamidronic acid, pegaspargase, peginterferonalfa-2b, pentosan polysulfate sodium, pentostatin, picibanil,pirarubicin, rabbit antithymocyte polyclonal antibody, polyethyleneglycol interferon alfa-2a, porfimer sodium, raloxifene, raltitrexed,rasburiembodiment, rhenium Re 186 etidronate, RII retinamide, rituximab,romurtide, samarium (153 Sm) lexidronam, sargramostim, sizofiran,sobuzoxane, sonermin, strontium-89 chloride, suramin, tasonermin,tazarotene, tegafur, temoporfin, temozolomide, teniposide,tetrachlorodecaoxide, thalidomide, thymalfasin, thyrotropin alfa,topotecan, toremifene, tositumomab-iodine 131, trastuzumab, treosulfan,tretinoin, trilostane, trimetrexate, triptorelin, tumor necrosis factoralpha, natural, ubenimex, bladder cancer vaccine, Maruyama vaccine,melanoma lysate vaccine, valrubicin, verteporfin, vinorelbine,virulizin, zinostatin stimalamer, orzoledronic acid; abarelix; AE 941(Aeterna), ambamustine, antisense oligonucleotide, bcl-2 (Genta), APC8015 (Dendreon), decitabine, dexaminoglutethimide, diaziquone, EL 532(Elan), EM 800 (Endorecherche), eniluracil, etanidazole, fenretinide,filgrastim SD01 (Amgen), fulvestrant, galocitabine, gastrin 17immunogen, HLA-B7 gene therapy (Vical), granulocyte macrophage colonystimulating factor, histamine dihydrochloride, ibritumomab tiuxetan,ilomastat, IM 862 (Cytran), interleukin-2, iproxifene, LDI 200(Milkhaus), leridistim, lintuzumab, CA 125 MAb (Biomira), cancer MAb(Japan Pharmaceutical Development), HER-2 and Fc MAb (Medarex),idiotypic 105AD7 MAb (CRC Technology), idiotypic CEA MAb (Trilex),LYM-1-iodine 131 MAb (Techni clone), polymorphic epithelialmucin-yttrium 90 MAb (Antisoma), marimastat, menogaril, mitumomab,motexafin gadolinium, MX 6 (Galderma), nelarabine, nolatrexed, P 30protein, pegvisomant, pemetrexed, porfiromycin, prinomastat, RL 0903(Shire), rubitecan, satraplatin, sodium phenylacetate, sparfosic acid,SRL 172 (SR Pharma), SU 5416 (SUGEN), TA 077 (Tanabe),tetrathiomolybdate, thaliblastine, thrombopoietin, tin ethyletiopurpurin, tirapazamine, cancer vaccine (Biomira), melanoma vaccine(New York University), melanoma vaccine (Sloan Kettering Institute),melanoma oncolysate vaccine (New York Medical College), viral melanomacell lysates vaccine (Royal Newcastle Hospital), or valspodar.

Additional examples of therapeutic agents that may be used incombination with compounds of the invention include ipilimumab(Yervoy®); tremelimumab; galiximab; nivolumab, also known as BMS-936558(Opdivo®); pembrolizumab (Keytruda®); avelumab (Bavencio®); AMP224;BMS-936559; MPDL3280A, also known as RG7446; MEDI-570; AMG557; MGA271;IMP321; BMS-663513; PF-05082566; CDX-1127; anti-OX40 (Providence HealthServices); huMAbOX40L; atacicept; CP-870893; lucatumumab; dacetuzumab;muromonab-CD3; ipilumumab; MEDI4736 (Imfinzi®); MSB0010718C; AMP 224;adalimumab (Humira®); ado-trastuzumab emtansine (Kadcyla®); aflibercept(Eylea®); alemtuzumab (Campath®); basiliximab (Simulect®); belimumab(Benlysta®); basiliximab (Simulect®); belimumab (Benlysta®); brentuximabvedotin (Adcetris®); canakinumab (Maris®); certolizumab pegol (Cimzia®);daclizumab (Zenapax®); daratumumab (Darzalex®); denosumab (Prolia®);eculizumab (Soliris®); efalizumab (Raptiva®); gemtuzumab ozogamicin(Mylotarg®); golimumab (Simponi®); ibritumomab tiuxetan (Zevalin®);infliximab (Remicade®); motavizumab (Numax®); natalizumab (Tysabri®);obinutuzumab (Gazyva®); ofatumumab (Arzerra®); omalizumab (Xolair®);palivizumab (Synagis®); pertuzumab (Perjeta®); pertuzumab (Perjeta®);ranibizumab (Lucentis®); raxibacumab (Abthrax®); tocilizumab (Actemra®);tositumomab; tositumomab-i-131; tositumomab and tositumomab-i-131(Bexxar®); ustekinumab (Stelara®); AMG 102; AMG 386; AMG 479; AMG 655;AMG 706; AMG 745; and AMG 951.

The compounds described herein can be used in combination with theagents disclosed herein or other suitable agents, depending on thecondition being treated. Hence, in some embodiments the one or morecompounds of the disclosure will be co-administered with other therapiesas described herein. When used in combination therapy, the compoundsdescribed herein may be administered with the second agentsimultaneously or separately. This administration in combination caninclude simultaneous administration of the two agents in the same dosageform, simultaneous administration in separate dosage forms, and separateadministration. That is, a compound described herein and any of theagents described herein can be formulated together in the same dosageform and administered simultaneously. Alternatively, a compound of theinvention and any of the therapies described herein can besimultaneously administered, wherein both the agents are present inseparate formulations. In another alternative, a compound of the presentdisclosure can be administered and followed by any of the therapiesdescribed herein, or vice versa. In some embodiments of the separateadministration protocol, a compound of the invention and any of thetherapies described herein are administered a few minutes apart, or afew hours apart, or a few days apart.

In some embodiments of any of the methods described herein, the firsttherapy (e.g., a compound of the invention) and one or more additionaltherapies are administered simultaneously or sequentially, in eitherorder. The first therapeutic agent may be administered immediately, upto 1 hour, up to 2 hours, up to 3 hours, up to 4 hours, up to 5 hours,up to 6 hours, up to 7 hours, up to, 8 hours, up to 9 hours, up tohours, up to 11 hours, up to 12 hours, up to 13 hours, 14 hours, up tohours 16, up to 17 hours, up 18 hours, up to 19 hours up to 20 hours, upto 21 hours, up to 22 hours, up to 23 hours, up to 24 hours, or up to1-7, 1-14, 1-21 or 1-30 days before or after the one or more additionaltherapies.

The invention also features kits including (a) a pharmaceuticalcomposition including an agent (e.g., a compound of the invention)described herein, and (b) a package insert with instructions to performany of the methods described herein. In some embodiments, the kitincludes (a) a pharmaceutical composition including an agent (e.g., acompound of the invention) described herein, (b) one or more additionaltherapies (e.g., non-drug treatment or therapeutic agent), and (c) apackage insert with instructions to perform any of the methods describedherein.

As one aspect of the present invention contemplates the treatment of thedisease or symptoms associated therewith with a combination ofpharmaceutically active compounds that may be administered separately,the invention further relates to combining separate pharmaceuticalcompositions in kit form. The kit may comprise two separatepharmaceutical compositions: a compound of the present invention, andone or more additional therapies. The kit may comprise a container forcontaining the separate compositions such as a divided bottle or adivided foil packet. Additional examples of containers include syringes,boxes, and bags. In some embodiments, the kit may comprise directionsfor the use of the separate components. The kit form is particularlyadvantageous when the separate components are preferably administered indifferent dosage forms (e.g., oral and parenteral), are administered atdifferent dosage intervals, or when titration of the individualcomponents of the combination is desired by the prescribing health careprofessional.

Numbered Embodiments

[1] A compound, or pharmaceutically acceptable salt thereof, having thestructure of Formula Ia:

wherein A is optionally substituted 3 to 6-membered cycloalkylene,optionally substituted 3 to 6-membered heterocycloalkylene, optionallysubstituted 6-membered arylene, optionally substituted 5 to 6-memberedheteroarylene, optionally substituted C₂-C₄ alkylene, or optionallysubstituted C₂-C₄ alkenylene;

Y is

W is hydrogen, C₁-C₄ alkyl, optionally substituted C₁-C₃ heteroalkyl,optionally substituted 3 to 10-membered heterocycloalkyl, optionallysubstituted 3 to 10-membered cycloalkyl, optionally substituted 6 to10-membered aryl, or optionally substituted 5 to 10-membered heteroaryl;

X¹ and X⁴ are each, independently, CH2 or NH;

R¹ is optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 15-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl;

R² is hydrogen, optionally substituted C₁-C₆ alkyl, optionallysubstituted C₂-C₆ alkenyl, optionally substituted C₂-C₆ alkynyl,optionally substituted 3 to 6-membered cycloalkyl, optionallysubstituted 3 to 7-membered heterocycloalkyl, optionally substituted6-membered aryl, optionally substituted 5 or 6-membered heteroaryl; and

R¹⁰ is hydrogen, hydroxy, optionally substituted C₁-C₃ alkyl, oroptionally substituted C₁-C₆ heteroalkyl.

[2] The compound of paragraph [1], or pharmaceutically acceptable saltthereof, wherein R¹ is optionally substituted 6 to 10-membered aryl oroptionally substituted 5 to 10-membered heteroaryl.

[3] The compound of paragraph [2], or pharmaceutically acceptable saltthereof, wherein R¹ is optionally substituted phenyl or optionallysubstituted pyridine.

[4] The compound of any one of paragraphs [1]-[3], or pharmaceuticallyacceptable salt thereof, wherein A is optionally substituted thiazole,optionally substituted triazole, optionally substituted morpholino,optionally substituted piperidinyl, optionally substituted pyridine, oroptionally substituted phenyl.

[5] The compound of any one of paragraphs [1]-[3], or pharmaceuticallyacceptable salt thereof, wherein A is not an optionally substitutedphenyl or benzimidazole.

[6] The compound of paragraph 5, or pharmaceutically acceptable saltthereof, wherein A is not hydroxyphenyl.

[7] The compound of any one of paragraphs [1]-[6], or pharmaceuticallyacceptable salt thereof, wherein the compound is not a compound of Table2.

[8] The compound of any one of paragraphs [1]-[7], or pharmaceuticallyacceptable salt thereof, wherein the compound is not a compound of Table3.

[9] The compound of any one of paragraphs [1]-[8], or pharmaceuticallyacceptable salt thereof, wherein Y is —NHC(O)— or —NHC(O)NH—.

[10] The compound of paragraph [9] or pharmaceutically acceptable saltthereof, having the structure of Formula IIa:

wherein a is 0 or 1.

[11] The compound of paragraph [10], or pharmaceutically acceptable saltthereof, having the structure of Formula II-1a:

wherein X² is N or CH;

each R³ is independently selected from halogen, cyano, hydroxy,optionally substituted amine, optionally substituted amido, optionallysubstituted C₁-C₆ alkyl, optionally substituted C₁-C₆ heteroalkyl,optionally substituted 3 to 6-membered cycloalkyl, optionallysubstituted 3 to 6-membered cycloalkenyl, optionally substituted 3 to11-membered heterocycloalkyl, optionally substituted 6 to 10-memberedaryl, or optionally substituted 5 to 10-membered heteroaryl; and

n is an integer from 1 to 4.

[12] The compound of paragraph [11], or pharmaceutically acceptable saltthereof, having the structure of Formula IIa-2:

[13] The compound of paragraph [12], or pharmaceutically acceptable saltthereof, having the structure of Formula IIa-3:

wherein R⁴ and R⁵ are each independently selected from halogen, cyano,hydroxy, optionally substituted amine, optionally substituted amido,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 11-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl.

[14] The compound of paragraph [13] or pharmaceutically acceptable saltthereof, having the structure of Formula IIa-4:

The compound of paragraph [14] or pharmaceutically acceptable saltthereof, having the structure of Formula IIa-5:

wherein X³ is N or CH;

m is 1 or 2;

R⁶, R⁷, R⁸, and R¹¹ are each independently selected from hydrogen,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 6-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl; or

R⁶ and R⁷ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered cycloalkyl or an optionallysubstituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R⁸ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R¹¹ combine with the atoms to which they are attached to form anoptionally substituted 4 to 8-membered heterocycloalkyl.

[16] The compound of paragraph [15], or pharmaceutically acceptable saltthereof, having the structure of Formula IIa-6:

[17] The compound of paragraph [15], or pharmaceutically acceptable saltthereof, having the structure of Formula IIa-7:

[18] The compound of paragraph [16] or [17], wherein R⁶ is methyl.

[19] The compound of paragraph [15], or pharmaceutically acceptable saltthereof, having the structure of Formula IIa-8 or Formula IIa-9:

[20] The compound of paragraph [9], or pharmaceutically acceptable saltthereof, having the structure of Formula IIIa:

wherein a is 0 or 1.

[21] The compound of paragraph [20], or pharmaceutically acceptable saltthereof, having the structure of Formula IIIa-1

wherein X² is N or CH;

each R³ is independently selected from halogen, cyano, hydroxy,optionally substituted amine, optionally substituted amido, optionallysubstituted C₁-C₆ alkyl, optionally substituted C₁-C₆ heteroalkyl,optionally substituted 3 to 6-membered cycloalkyl, optionallysubstituted 3 to 6-membered cycloalkenyl, optionally substituted 3 to11-membered heterocycloalkyl, optionally substituted 6 to 10-memberedaryl, or optionally substituted 5 to 10-membered heteroaryl; and

n is an integer from 1 to 4.

[22] The compound of paragraph [21], or pharmaceutically acceptable saltthereof, having the structure of Formula IIIa-2:

[23], The compound of paragraph [22], or pharmaceutically acceptablesalt thereof, having the structure of Formula IIIa-3:

wherein R⁴ and R⁵ are each independently selected from halogen, cyano,hydroxy, optionally substituted amine, optionally substituted amido,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 11-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl.

[24] The compound of paragraph [23], or pharmaceutically acceptable saltthereof, having the structure of Formula IIIa-4:

The compound of paragraph [24], or pharmaceutically acceptable saltthereof, having the structure of Formula IIIa-5:

wherein X³ is N or CH;

m is 1 or 2;

R⁶, R⁷, R⁸, and R¹¹ are each independently selected from hydrogen,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 6-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl; or

R⁶ and R⁷ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered cycloalkyl or an optionallysubstituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R⁸ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R¹¹ combine with the atoms to which they are attached to form anoptionally substituted 4 to 8-membered heterocycloalkyl.

[26] The compound of paragraph [25], or pharmaceutically acceptable saltthereof, having the structure of Formula IIIa-6:

[27] The compound of paragraph [25], or pharmaceutically acceptable saltthereof, having the structure of Formula IIIa-7:

[28] The compound of paragraph [26] or [27], wherein R⁶ is methyl.

[29] The compound of paragraph [25], or pharmaceutically acceptable saltthereof, having the structure of Formula IIIa-8 or Formula IIIa-9:

[30] The compound of paragraph [9], or pharmaceutically acceptable saltthereof, having the structure of Formula IVa:

wherein R⁹ is H or C₁-C₆ alkyl; and

a is 0 or 1.

[31] The compound of paragraph [30], or pharmaceutically acceptable saltthereof, having the structure of Formula IVa-1:

wherein X² is N or CH;

each R³ is independently selected from halogen, cyano, hydroxy,optionally substituted amine, optionally substituted amido, optionallysubstituted C₁-C₆ alkyl, optionally substituted C₁-C₆ heteroalkyl,optionally substituted 3 to 6-membered cycloalkyl, optionallysubstituted 3 to 6-membered cycloalkenyl, optionally substituted 3 to11-membered heterocycloalkyl, optionally substituted 6 to 10-memberedaryl, or optionally substituted 5 to 10-membered heteroaryl; and

n is an integer from 1 to 4.

[32] The compound of paragraph [31], or pharmaceutically acceptable saltthereof, having the structure of Formula IVa-2:

[33] The compound of paragraph [32], or pharmaceutically acceptable saltthereof, having the structure of Formula IVa-3:

wherein R⁴ and R⁵ are each independently selected from halogen, cyano,hydroxy, optionally substituted amine, optionally substituted amido,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 11-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl.

[34] The compound of paragraph [33], or pharmaceutically acceptable saltthereof, having the structure of Formula IVa-4:

[35] The compound of paragraph [34], or pharmaceutically acceptable saltthereof, having the structure of Formula IVa-5:

wherein X³ is N or CH;

m is 1 or 2;

R⁶, R⁷, R⁸, and R¹¹ are each independently selected from hydrogen,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 6-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl; or

R⁶ and R⁷ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered cycloalkyl or an optionallysubstituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R⁸ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R¹¹ combine with the atoms to which they are attached to form anoptionally substituted 4 to 8-membered heterocycloalkyl.

[36] The compound of paragraph [35], or pharmaceutically acceptable saltthereof, having the structure of Formula IVa-6:

[37] The compound of paragraph [35], or pharmaceutically acceptable saltthereof, having the structure of Formula IVa-7:

[38] The compound of paragraph [36] or [37], wherein R⁶ is methyl.

[39] The compound of paragraph [35], or pharmaceutically acceptable saltthereof, having the structure of Formula IVa-8 or Formula IVa-9:

[40] The compound of any one of paragraphs [30]-[39], orpharmaceutically acceptable salt thereof, wherein R⁹ is methyl.

[41] The compound of any one of paragraphs [1]-[8], or pharmaceuticallyacceptable salt thereof, wherein Y is —NHS(O)₂— or —NHS(O)₂NH—.

[42] The compound of paragraph [41], or pharmaceutically acceptable saltthereof, having the structure of Formula Va:

wherein a is 0 or 1.

[43] The compound of paragraph [42], or pharmaceutically acceptable saltthereof, having the structure of Formula Va-1:

wherein X² is N or CH;

each R³ is independently selected from halogen, cyano, hydroxy,optionally substituted amine, optionally substituted amido, optionallysubstituted C₁-C₆ alkyl, optionally substituted C₁-C₆ heteroalkyl,optionally substituted 3 to 6-membered cycloalkyl, optionallysubstituted 3 to 6-membered cycloalkenyl, optionally substituted 3 to11-membered heterocycloalkyl, optionally substituted 6 to 10-memberedaryl, or optionally substituted 5 to 10-membered heteroaryl; and

n is an integer from 1 to 4.

[44] The compound of paragraph [43], or pharmaceutically acceptable saltthereof, having the structure of Formula Va-2:

[45] The compound of paragraph [44], or pharmaceutically acceptable saltthereof, having the structure of Formula Va-3:

wherein R⁴ and R⁵ are each independently selected from halogen, cyano,hydroxy, optionally substituted amine, optionally substituted amido,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 11-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl.

[46] The compound of paragraph [45], or pharmaceutically acceptable saltthereof, having the structure of Formula Va-4:

[47] The compound of paragraph [46], or pharmaceutically acceptable saltthereof, having the structure of Formula Va-5:

wherein X³ is N or CH;

m is 1 or 2;

R⁶, R⁷, R⁸, and R¹¹ are each independently selected from hydrogen,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 6-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl; or

R⁶ and R⁷ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered cycloalkyl or an optionallysubstituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R⁸ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R¹¹ combine with the atoms to which they are attached to form anoptionally substituted 4 to 8-membered heterocycloalkyl.

[48] The compound of paragraph [41], or pharmaceutically acceptable saltthereof, having the structure of Formula VIa:

wherein a is 0 or 1.

[49] The compound of paragraph [48], or pharmaceutically acceptable saltthereof, having the structure of Formula VIa-1:

wherein X² is N or CH;

each R³ is independently selected from halogen, cyano, hydroxy,optionally substituted amine, optionally substituted amido, optionallysubstituted C₁-C₆ alkyl, optionally substituted C₁-C₆ heteroalkyl,optionally substituted 3 to 6-membered cycloalkyl, optionallysubstituted 3 to 6-membered cycloalkenyl, optionally substituted 3 to11-membered heterocycloalkyl, optionally substituted 6 to 10-memberedaryl, or optionally substituted 5 to 10-membered heteroaryl; and

n is an integer from 1 to 4.

[50] The compound of paragraph [49], or pharmaceutically acceptable saltthereof, having the structure of Formula VIa-2:

[51] The compound of paragraph [50], or pharmaceutically acceptable saltthereof, having the structure of Formula VIa-3:

wherein R⁴ and R⁵ are each independently selected from halogen, cyano,hydroxy, optionally substituted amine, optionally substituted amido,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 11-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl.

[52] The compound of paragraph [51], or pharmaceutically acceptable saltthereof, having the structure of Formula VIa-4:

[53] The compound of paragraph [52], or pharmaceutically acceptable saltthereof, having the structure of Formula VIa-5:

wherein X³ is N or CH;

m is 1 or 2;

R⁶, R⁷, R⁸, and R¹¹ are each independently selected from hydrogen,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 6-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl; or

R⁶ and R⁷ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered cycloalkyl or an optionallysubstituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R⁸ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R¹¹ combine with the atoms to which they are attached to form anoptionally substituted 4 to 8-membered heterocycloalkyl.

[54] The compound of paragraph [41], or pharmaceutically acceptable saltthereof, having the structure of Formula VIIa:

wherein R⁹ is H or C₁-C₆ alkyl; and

a is 0 or 1.

[55] The compound of paragraph [54], or pharmaceutically acceptable saltthereof, having the structure of Formula VIIa-1:

wherein X² is N or CH;

each R³ is independently selected from halogen, cyano, hydroxy,optionally substituted amine, optionally substituted amido, optionallysubstituted C₁-C₆ alkyl, optionally substituted C₁-C₆ heteroalkyl,optionally substituted 3 to 6-membered cycloalkyl, optionallysubstituted 3 to 6-membered cycloalkenyl, optionally substituted 3 to11-membered heterocycloalkyl, optionally substituted 6 to 10-memberedaryl, or optionally substituted 5 to 10-membered heteroaryl; and

n is an integer from 1 to 4.

[56] The compound of paragraph [55], or pharmaceutically acceptable saltthereof, having the structure of Formula VIIa-2:

[57] The compound of paragraph [56], or pharmaceutically acceptable saltthereof, having the structure of Formula VIIa-3:

wherein R⁴ and R⁵ are each independently selected from halogen, cyano,hydroxy, optionally substituted amine, optionally substituted amido,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 11-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl.

[58] The compound of paragraph [57], or pharmaceutically acceptable saltthereof, having the structure of Formula VIIa-4:

[59] The compound of paragraph [58], or pharmaceutically acceptable saltthereof, having the structure of Formula VIIa-5:

wherein X³ is N or CH;

m is 1 or 2;

R⁶, R⁷, R⁸, and R¹¹ are each independently selected from hydrogen,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 6-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl; or

R⁶ and R⁷ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered cycloalkyl or an optionallysubstituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R⁸ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R¹¹ combine with the atoms to which they are attached to form anoptionally substituted 4 to 8-membered heterocycloalkyl.

[60] The compound of any one of paragraphs [54]-[59], orpharmaceutically acceptable salt thereof, wherein R⁹ is methyl.

[61] The compound of any one of paragraphs [1]-[8], or pharmaceuticallyacceptable salt thereof, wherein Y is —NHS(O)— or —NHS(O)NH—.

[62] The compound of paragraph [61], or pharmaceutically acceptable saltthereof, having the structure of Formula VIIIa:

wherein a is 0 or 1.

[63] The compound of paragraph [62], or pharmaceutically acceptable saltthereof, having the structure of Formula VIIIa-1:

wherein X² is N or CH;

each R³ is independently selected from halogen, cyano, hydroxy,optionally substituted amine, optionally substituted amido, optionallysubstituted C₁-C₆ alkyl, optionally substituted C₁-C₆ heteroalkyl,optionally substituted 3 to 6-membered cycloalkyl, optionallysubstituted 3 to 6-membered cycloalkenyl, optionally substituted 3 to11-membered heterocycloalkyl, optionally substituted 6 to 10-memberedaryl, or optionally substituted 5 to 10-membered heteroaryl; and

n is an integer from 1 to 4.

[64] The compound of paragraph [63], or pharmaceutically acceptable saltthereof, having the structure of Formula VIIIa-2:

[65] The compound of paragraph [64], or pharmaceutically acceptable saltthereof, having the structure of Formula VIIIa-3:

wherein R⁴ and R⁵ are each independently selected from halogen, cyano,hydroxy, optionally substituted amine, optionally substituted amido,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 11-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl.

[66] The compound of paragraph [65], or pharmaceutically acceptable saltthereof, having the structure of Formula VIIIa-4:

[67] The compound of paragraph [66], or pharmaceutically acceptable saltthereof, having the structure of Formula VIIIa-5:

wherein X³ is N or CH;

m is 1 or 2;

R⁶, R⁷, R⁸, and R¹¹ are each independently selected from hydrogen,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 6-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl; or

R⁶ and R⁷ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered cycloalkyl or an optionallysubstituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R⁸ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R¹¹ combine with the atoms to which they are attached to form anoptionally substituted 4 to 8-membered heterocycloalkyl.

[68] The compound of paragraph [61], or pharmaceutically acceptable saltthereof, having the structure of Formula IXa:

wherein a is 0 or 1.

[69] The compound of paragraph [68], or pharmaceutically acceptable saltthereof, having the structure of Formula IXa-1:

wherein X² is N or CH;

each R³ is independently selected from halogen, cyano, hydroxy,optionally substituted amine, optionally substituted amido, optionallysubstituted C₁-C₆ alkyl, optionally substituted C₁-C₆ heteroalkyl,optionally substituted 3 to 6-membered cycloalkyl, optionallysubstituted 3 to 6-membered cycloalkenyl, optionally substituted 3 to11-membered heterocycloalkyl, optionally substituted 6 to 10-memberedaryl, or optionally substituted 5 to 10-membered heteroaryl; and

n is an integer from 1 to 4.

[70] The compound of paragraph [69], or pharmaceutically acceptable saltthereof, having the structure of Formula IXa-2:

[71] The compound of paragraph [70], or pharmaceutically acceptable saltthereof, having the structure of Formula IXa-3:

wherein R⁴ and R⁵ are each independently selected from halogen, cyano,hydroxy, optionally substituted amine, optionally substituted amido,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 11-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl.

The compound of paragraph [71], or pharmaceutically acceptable saltthereof, having the structure of Formula IXa-4:

[73] The compound of paragraph [72], or pharmaceutically acceptable saltthereof, having the structure of Formula IXa-5:

wherein X³ is N or CH;

m is 1 or 2;

R⁶, R⁷, R⁸, and R¹¹ are each independently selected from hydrogen,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 6-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl; or

R⁶ and R⁷ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered cycloalkyl or an optionallysubstituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R⁸ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R¹¹ combine with the atoms to which they are attached to form anoptionally substituted 4 to 8-membered heterocycloalkyl.

[74] The compound of paragraph [61], or pharmaceutically acceptable saltthereof, having the structure of Formula Xa:

wherein R⁹ is H or C₁-C₆ alkyl; and

a is 0 or 1.

[75] The compound of paragraph [74], or pharmaceutically acceptable saltthereof, having the structure of Formula Xa-1:

wherein X² is N or CH;

each R³ is independently selected from halogen, cyano, hydroxy,optionally substituted amine, optionally substituted amido, optionallysubstituted C₁-C₆ alkyl, optionally substituted C₁-C₆ heteroalkyl,optionally substituted 3 to 6-membered cycloalkyl, optionallysubstituted 3 to 6-membered cycloalkenyl, optionally substituted 3 to11-membered heterocycloalkyl, optionally substituted 6 to 10-memberedaryl, or optionally substituted 5 to 10-membered heteroaryl; and

n is an integer from 1 to 4.

[76] The compound of paragraph [75], or pharmaceutically acceptable saltthereof, having the structure of Formula Xa-2:

[77] The compound of paragraph [76], or pharmaceutically acceptable saltthereof, having the structure of Formula Xa-3:

wherein R⁴ and R⁵ are each independently selected from halogen, cyano,hydroxy, optionally substituted amine, optionally substituted amido,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 11-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl.

[78] The compound of paragraph [77] or pharmaceutically acceptable saltthereof, having the structure of Formula Xa-4:

[79] The compound of paragraph [78], or pharmaceutically acceptable saltthereof, having the structure of Formula Xa-5:

wherein X³ is N or CH;

m is 1 or 2;

R⁶, R⁷, R⁸, and R¹¹ are each independently selected from hydrogen,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 6-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl; or

R⁶ and R⁷ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered cycloalkyl or an optionallysubstituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R⁸ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R¹¹ combine with the atoms to which they are attached to form anoptionally substituted 4 to 8-membered heterocycloalkyl.

[80] The compound of any one of paragraphs [74]-[79], orpharmaceutically acceptable salt thereof, wherein R⁹ is methyl.

[81] The compound of any one of paragraphs [10]-[40], [42]-[60], or[62]-[80], or pharmaceutically acceptable salt thereof, wherein a is 0.

[82] The compound of any one of paragraphs [10]-[40], [42]-[60], or[62]-[80], or pharmaceutically acceptable salt thereof, wherein a is 1.

[83] The compound of any one of paragraphs [1]-[82], or pharmaceuticallyacceptable salt thereof, wherein R² is optionally substituted C₁-C₆alkyl.

[84] The compound of paragraph [83], or pharmaceutically acceptable saltthereof, wherein R² is selected from —CH₂CH₃ or—CH₂CF₃.

[85] The compound of any one of paragraphs [1]-[84], or pharmaceuticallyacceptable salt thereof, wherein W is C₁-C₄ alkyl.

[86] The compound of any one of paragraphs [1]-[84], or pharmaceuticallyacceptable salt thereof, wherein W is optionally substitutedcyclopropyl, optionally substituted cyclobutyl, optionally substitutedcyclopentyl, or optionally substituted cyclohexyl, optionallysubstituted piperidine, optionally substituted piperazine, optionallysubstituted pyridine, or optionally substituted phenyl.

[87] The compound of any one of paragraphs [1]-[84], or pharmaceuticallyacceptable salt thereof, wherein W is optionally substituted 3 to10-membered heterocycloalkyl, optionally substituted 3 to 10-memberedcycloalkyl, optionally substituted 6 to 10-membered aryl, or optionallysubstituted 5 to 10-membered heteroaryl.

[88] The compound of any one of paragraphs [1]-[84], or pharmaceuticallyacceptable salt thereof, wherein W is optionally substituted 3 to10-membered heterocycloalkyl.

[89] The compound of paragraph [88], or pharmaceutically acceptable saltthereof, wherein W is selected from the following, or a stereoisomerthereof:

[90] The compound of any one of paragraphs [1]-[84], or pharmaceuticallyacceptable salt thereof, wherein W is optionally substituted 3 to10-membered cycloalkyl.

[91] The compound of paragraph [90], or pharmaceutically acceptable saltthereof, wherein W is selected from the following, or a stereoisomerthereof:

[92] The compound of any one of paragraphs [1]-[84], or pharmaceuticallyacceptable salt thereof, wherein W is optionally substituted 5 to10-membered heteroaryl.

[93] The compound of paragraph [92], or pharmaceutically acceptable saltthereof, wherein W is selected from the following, or a stereoisomerthereof:

[94] The compound of any one of paragraphs [1]-[84], or pharmaceuticallyacceptable salt thereof, wherein W is optionally substituted 6 to10-membered aryl.

[95] The compound of paragraph [94], or pharmaceutically acceptable saltthereof, wherein W is optionally substituted phenyl.

[96] The compound of any one of paragraphs [1]-[84], or pharmaceuticallyacceptable salt thereof, wherein W is optionally substituted C₁-C₃heteroalkyl.

[97] The compound of paragraph [96], or pharmaceutically acceptable saltthereof, wherein W is selected from the following, or a stereoisomerthereof:

[98] The compound of paragraph [85], or pharmaceutically acceptable saltthereof, wherein W is selected from the following:

[99] A compound, or a pharmaceutically acceptable salt thereof, of Table1a.

[100] A compound, or a pharmaceutically acceptable salt thereof, ofTable 1b.

[101] A compound, or pharmaceutically acceptable salt thereof, havingthe structure of Formula

wherein A is optionally substituted 3 to 6-membered cycloalkylene,optionally substituted 3 to 6-membered heterocycloalkylene, optionallysubstituted 6-membered arylene, optionally substituted 5 to 6-memberedheteroarylene, optionally substituted C₂-C₄ alkylene, or optionallysubstituted C₂-C₄ alkenylene;

Y is

W is hydrogen, C₁-C₄ alkyl, optionally substituted C₁-C₃ heteroalkyl,optionally substituted 3 to 10-membered heterocycloalkyl, optionallysubstituted 3 to 10-membered cycloalkyl, optionally substituted 6 to10-membered aryl, or optionally substituted 5 to 10-membered heteroaryl;or W is —R¹⁴C(═O)R¹⁵ where R¹⁴ is 3 to 10-membered cycloalkylene and R¹⁵is selected from optionally substituted 3 to 10-membered cycloalkyl,optionally substituted 6 to 10-membered aryl, or optionally substituted5 to 10-membered heteroaryl;

X¹ and X⁴ are each, independently, CH₂, CH(CH₃) or NH;

R¹ is optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 15-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl;

R² is hydrogen, optionally substituted C₁-C₆ alkyl, optionallysubstituted C₂-C₆ alkenyl, optionally substituted C₂-C₆ alkynyl,optionally substituted 3 to 6-membered cycloalkyl, optionallysubstituted 3 to 7-membered heterocycloalkyl, optionally substituted6-membered aryl, optionally substituted 5 or 6-membered heteroaryl;

R¹⁰ is hydrogen, hydroxy, optionally substituted C₁-C₆ alkoxy,optionally substituted C₁-C₃ alkyl, optionally substituted C₁-C₆heteroalkyl, or optionally substituted 3 to 7-membered heterocycloalkyl;and

R¹² and R¹³ are each, independently, selected from F or CH₃, or R¹² andR¹³ combine with the atoms to which they are attached to make a3-membered cycloalkyl.

[102] The compound of paragraph [101], or pharmaceutically acceptablesalt thereof, wherein R¹ is optionally substituted 6 to 10-membered arylor optionally substituted 5 to 10-membered heteroaryl.

[103] The compound of paragraph [102], or pharmaceutically acceptablesalt thereof, wherein R¹ is optionally substituted phenyl or optionallysubstituted pyridine.

[104] The compound of any one of paragraphs [101]-[103], orpharmaceutically acceptable salt thereof, wherein A is optionallysubstituted thiazole, optionally substituted triazole, optionallysubstituted morpholino, optionally substituted piperidinyl, optionallysubstituted pyridine, or optionally substituted phenyl.

[105] The compound of any one of paragraphs [101]-[103], orpharmaceutically acceptable salt thereof, wherein A is not an optionallysubstituted phenyl or benzimidazole.

[106] The compound of paragraph [105], or pharmaceutically acceptablesalt thereof, wherein A is not hydroxyphenyl.

[107] The compound of any one of paragraphs [101]-[106], orpharmaceutically acceptable salt thereof, wherein the compound is not acompound of Table 2.

[108] The compound of any one of paragraphs [101]-[107], orpharmaceutically acceptable salt thereof, wherein the compound is not acompound of Table 3.

[109] The compound of any one of paragraphs [101]-[108], orpharmaceutically acceptable salt thereof, wherein Y is —NHC(O)—or—NHC(O)NH—.

[110] The compound of paragraph [109], or pharmaceutically acceptablesalt thereof, having the structure of Formula IIb:

wherein a is 0 or 1.

[111] The compound of paragraph [110], or pharmaceutically acceptablesalt thereof, having the structure of Formula IIb-1:

wherein X² is N or CH;

each R³ is independently selected from halogen, cyano, hydroxy,optionally substituted amine, optionally substituted amido, optionallysubstituted C₁-C₆ alkyl, optionally substituted C₁-C₆ heteroalkyl,optionally substituted 3 to 6-membered cycloalkyl, optionallysubstituted 3 to 6-membered cycloalkenyl, optionally substituted 3 to11-membered heterocycloalkyl, optionally substituted 6 to 10-memberedaryl, or optionally substituted 5 to 10-membered heteroaryl; and

n is an integer from 1 to 4.

[112] The compound of paragraph [111], or pharmaceutically acceptablesalt thereof, having the structure of Formula IIb-2:

[113] The compound of paragraph [112], or pharmaceutically acceptablesalt thereof, having the structure of Formula IIb-3:

wherein R⁴ and R⁵ are each independently selected from hydrogen,halogen, cyano, hydroxy, optionally substituted amine, optionallysubstituted amido, optionally substituted C₁-C₆ alkyl, optionallysubstituted C₁-C₆ heteroalkyl, optionally substituted 3 to 6-memberedcycloalkyl, optionally substituted 3 to 6-membered cycloalkenyl,optionally substituted 3 to 11-membered heterocycloalkyl, optionallysubstituted 6 to 10-membered aryl, or optionally substituted 5 to10-membered heteroaryl. In some embodiments, R⁴ and R⁵ are not hydrogen.

In some embodiments, the compound, or pharmaceutically acceptable saltthereof, has the structure of Formula IIb-3, wherein W is optionallysubstituted 3 to 10-membered heterocycloalkyl, optionally substituted 3to 10-membered cycloalkyl, optionally substituted 6 to 10-membered aryl,or optionally substituted 5 to 10-membered heteroaryl; or W is—R¹⁴C(═O)R¹⁵ where R¹⁴ is 3 to 10-membered cycloalkyl and R¹⁵ isselected from optionally substituted 3 to 10-membered cycloalkyl,optionally substituted 6 to 10-membered aryl, or optionally substituted5 to 10-membered heteroaryl; R² is hydrogen, optionally substitutedC₁-C₆ alkyl, optionally substituted C₂-C₆ alkenyl, optionallysubstituted C₂-C₆ alkynyl, optionally substituted 3 to 6-memberedcycloalkyl, optionally substituted 3 to 7-membered heterocycloalkyl,optionally substituted 6-membered aryl, optionally substituted 5 or6-membered heteroaryl; R⁴ and R⁵ are each independently selected fromhalogen, cyano, hydroxy, optionally substituted amine, optionallysubstituted amido, optionally substituted C₁-C₆ alkyl, optionallysubstituted C₁-C₆ heteroalkyl, optionally substituted 3 to 6-memberedcycloalkyl, optionally substituted 3 to 6-membered cycloalkenyl,optionally substituted 3 to 11-membered heterocycloalkyl, optionallysubstituted 6 to 10-membered aryl, or optionally substituted 5 to10-membered heteroaryl; and R¹⁰ is hydrogen, hydroxy, optionallysubstituted C₁-C₆ alkoxy, optionally substituted C₁-C₃ alkyl, optionallysubstituted C₁-C₆ heteroalkyl, or optionally substituted 3 to 7-memberedheterocycloalkyl.

[114] The compound of paragraph [113], or pharmaceutically acceptablesalt thereof, having the structure of Formula IIb-4:

In some embodiments, the compound, or pharmaceutically acceptable saltthereof, has the structure of Formula IIb-4, wherein W is optionallysubstituted 3 to 10-membered heterocycloalkyl, optionally substituted 3to 10-membered cycloalkyl, optionally substituted 6 to 10-membered aryl,or optionally substituted 5 to 10-membered heteroaryl; or W is—R¹⁴C(═O)R¹⁵ where R¹⁴ is 3 to 10-membered cycloalkyl and R¹⁵ isselected from optionally substituted 3 to 10-membered cycloalkyl,optionally substituted 6 to 10-membered aryl, or optionally substituted5 to 10-membered heteroaryl; R² is hydrogen, optionally substitutedC₁-C₆ alkyl, optionally substituted C₂-C₆ alkenyl, optionallysubstituted C₂-C₆ alkynyl, optionally substituted 3 to 6-memberedcycloalkyl, optionally substituted 3 to 7-membered heterocycloalkyl,optionally substituted 6-membered aryl, optionally substituted 5 or6-membered heteroaryl; R⁵ is selected from halogen, cyano, hydroxy,optionally substituted amine, optionally substituted amido, optionallysubstituted C₁-C₆ alkyl, optionally substituted C₁-C₆ heteroalkyl,optionally substituted 3 to 6-membered cycloalkyl, optionallysubstituted 3 to 6-membered cycloalkenyl, optionally substituted 3 to11-membered heterocycloalkyl, optionally substituted 6 to 10-memberedaryl, or optionally substituted 5 to 10-membered heteroaryl; and R¹⁰ ishydrogen, hydroxy, optionally substituted C₁-C₆ alkoxy, optionallysubstituted C₁-C₃ alkyl, optionally substituted C₁-C₆ heteroalkyl, oroptionally substituted 3 to 7-membered heterocycloalkyl.

[115] The compound of paragraph [114], or pharmaceutically acceptablesalt thereof, having the structure of Formula IIb-5:

wherein X³ is N or CH;

m is 1 or 2;

R⁶, R⁷, R⁸, and R¹¹ are each independently selected from hydrogen,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 6-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl; or

R⁶ and R⁷ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered cycloalkyl or an optionallysubstituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R⁸ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R¹¹ combine with the atoms to which they are attached to form anoptionally substituted 4 to 8-membered heterocycloalkyl.

[116] The compound of paragraph [115], or pharmaceutically acceptablesalt thereof, having the structure of Formula IIb-6:

[117] The compound of paragraph [115], or pharmaceutically acceptablesalt thereof, having the structure of Formula IIb-7:

[118] The compound of paragraph [116] or [117], wherein R⁶ is methyl.

[119] The compound of paragraph [115], or pharmaceutically acceptablesalt thereof, having the structure of Formula IIb-8 or Formula IIb-9:

[120] The compound of paragraph [109], or pharmaceutically acceptablesalt thereof, having the structure of Formula IIIb:

wherein a is 0 or 1.

[121] The compound of paragraph [120], or pharmaceutically acceptablesalt thereof, having the structure of Formula IIIb-1:

wherein X² is N or CH;

each R³ is independently selected from halogen, cyano, hydroxy,optionally substituted amine, optionally substituted amido, optionallysubstituted C₁-C₆ alkyl, optionally substituted C₁-C₆ heteroalkyl,optionally substituted 3 to 6-membered cycloalkyl, optionallysubstituted 3 to 6-membered cycloalkenyl, optionally substituted 3 to11-membered heterocycloalkyl, optionally substituted 6 to 10-memberedaryl, or optionally substituted 5 to 10-membered heteroaryl; and

n is an integer from 1 to 4.

[122] The compound of paragraph [121], or pharmaceutically acceptablesalt thereof, having the structure of Formula IIIb-2:

[123] The compound of paragraph [122], or pharmaceutically acceptablesalt thereof, having the structure of Formula IIIb-3:

wherein R⁴ and R⁵ are each independently selected from hydrogen,halogen, cyano, hydroxy, optionally substituted amine, optionallysubstituted amido, optionally substituted C₁-C₆ alkyl, optionallysubstituted C₁-C₆ heteroalkyl, optionally substituted 3 to 6-memberedcycloalkyl, optionally substituted 3 to 6-membered cycloalkenyl,optionally substituted 3 to 11-membered heterocycloalkyl, optionallysubstituted 6 to 10-membered aryl, or optionally substituted 5 to10-membered heteroaryl. In some embodiments, R⁴ and R⁵ are not hydrogen.

[124] The compound of paragraph [123], or pharmaceutically acceptablesalt thereof, having the structure of Formula IIIb-4:

[125] The compound of paragraph [124], or pharmaceutically acceptablesalt thereof, having the structure of Formula IIIb-5:

wherein X³ is N or CH;

m is 1 or 2;

R⁶, R⁷, R⁸, and R¹¹ are each independently selected from hydrogen,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 6-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl; or

R⁶ and R⁷ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered cycloalkyl or an optionallysubstituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R⁸ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R¹¹ combine with the atoms to which they are attached to form anoptionally substituted 4 to 8-membered heterocycloalkyl.

[126] The compound of paragraph [125], or pharmaceutically acceptablesalt thereof, having the structure of Formula IIIb-6:

[127] The compound of paragraph [125], or pharmaceutically acceptablesalt thereof, having the structure of Formula IIIb-7:

[128] The compound of paragraph [126] or [127], wherein R⁶ is methyl.

[129] The compound of paragraph [125], or pharmaceutically acceptablesalt thereof, having the structure of Formula IIIb-8 or Formula IIIb-9:

[130] The compound of paragraph [109], or pharmaceutically acceptablesalt thereof, having the structure of Formula IVb:

wherein R⁹ is H or C₁-C₆ alkyl; and

a is 0 or 1.

[131] The compound of paragraph [130], or pharmaceutically acceptablesalt thereof, having the structure of Formula IVb-1:

wherein X² is N or CH;

each R³ is independently selected from halogen, cyano, hydroxy,optionally substituted amine, optionally substituted amido, optionallysubstituted C₁-C₆ alkyl, optionally substituted C₁-C₆ heteroalkyl,optionally substituted 3 to 6-membered cycloalkyl, optionallysubstituted 3 to 6-membered cycloalkenyl, optionally substituted 3 to11-membered heterocycloalkyl, optionally substituted 6 to 10-memberedaryl, or optionally substituted 5 to 10-membered heteroaryl; and

n is an integer from 1 to 4.

[132] The compound of paragraph [131], or pharmaceutically acceptablesalt thereof, having the structure of Formula IVb-2:

[133] The compound of paragraph [132], or pharmaceutically acceptablesalt thereof, having the structure of Formula IVb-3:

wherein R⁴ and R⁵ are each independently selected from hydrogen,halogen, cyano, hydroxy, optionally substituted amine, optionallysubstituted amido, optionally substituted C₁-C₆ alkyl, optionallysubstituted C₁-C₆ heteroalkyl, optionally substituted 3 to 6-memberedcycloalkyl, optionally substituted 3 to 6-membered cycloalkenyl,optionally substituted 3 to 11-membered heterocycloalkyl, optionallysubstituted 6 to 10-membered aryl, or optionally substituted 5 to10-membered heteroaryl. In some embodiments, R⁴ and R⁵ are not hydrogen.

[134] The compound of paragraph [133], or pharmaceutically acceptablesalt thereof, having the structure of Formula IVb-4:

[135] The compound of paragraph [134], or pharmaceutically acceptablesalt thereof, having the structure of Formula IVb-5:

wherein X³ is N or CH;

m is 1 or 2;

R⁶, R⁷, R⁸, and R¹¹ are each independently selected from hydrogen,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 6-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl; or

R⁶ and R⁷ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered cycloalkyl or an optionallysubstituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R⁸ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R¹¹ combine with the atoms to which they are attached to form anoptionally substituted 4 to 8-membered heterocycloalkyl.

[136] The compound of paragraph [135], or pharmaceutically acceptablesalt thereof, having the structure of Formula IVb-6:

[137] The compound of paragraph [135], or pharmaceutically acceptablesalt thereof, having the structure of Formula IVb-7:

[138] The compound of paragraph [136] or [137], wherein R⁶ is methyl.

[139] The compound of paragraph [135], or pharmaceutically acceptablesalt thereof, having the structure of Formula IVb-8 or Formula IVb-9:

[140] The compound of any one of paragraphs [130]-[139], orpharmaceutically acceptable salt thereof, wherein R⁹ is methyl.

[141] The compound of any one of paragraphs [101]-[108], orpharmaceutically acceptable salt thereof, wherein Y is —NHS(O)₂— or—NHS(O)₂NH—.

[142] The compound of paragraph [141], or pharmaceutically acceptablesalt thereof, having the structure of Formula Vb:

wherein a is 0 or 1.

[143] The compound of paragraph [142], or pharmaceutically acceptablesalt thereof, having the structure of Formula Vb-1:

wherein X² is N or CH;

each R³ is independently selected from halogen, cyano, hydroxy,optionally substituted amine, optionally substituted amido, optionallysubstituted C₁-C₆ alkyl, optionally substituted C₁-C₆ heteroalkyl,optionally substituted 3 to 6-membered cycloalkyl, optionallysubstituted 3 to 6-membered cycloalkenyl, optionally substituted 3 to11-membered heterocycloalkyl, optionally substituted 6 to 10-memberedaryl, or optionally substituted 5 to 10-membered heteroaryl; and

n is an integer from 1 to 4.

[144] The compound of paragraph [143], or pharmaceutically acceptablesalt thereof, having the structure of Formula Vb-2:

[145] The compound of paragraph [144], or pharmaceutically acceptablesalt thereof, having the structure of Formula Vb-3:

wherein R⁴ and R⁵ are each independently selected from hydrogen,halogen, cyano, hydroxy, optionally substituted amine, optionallysubstituted amido, optionally substituted C₁-C₆ alkyl, optionallysubstituted C₁-C₆ heteroalkyl, optionally substituted 3 to 6-memberedcycloalkyl, optionally substituted 3 to 6-membered cycloalkenyl,optionally substituted 3 to 11-membered heterocycloalkyl, optionallysubstituted 6 to 10-membered aryl, or optionally substituted 5 to10-membered heteroaryl. In some embodiments, R⁴ and R⁵ are not hydrogen.

[146] The compound of paragraph [145], or pharmaceutically acceptablesalt thereof, having the structure of Formula Vb-4:

[147] The compound of paragraph [146], or pharmaceutically acceptablesalt thereof, having the structure of Formula Vb-5:

wherein X³ is N or CH;

m is 1 or 2;

R⁶, R⁷, R⁸, and R¹¹ are each independently selected from hydrogen,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 6-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl; or

R⁶ and R⁷ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered cycloalkyl or an optionallysubstituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R⁸ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R¹¹ combine with the atoms to which they are attached to form anoptionally substituted 4 to 8-membered heterocycloalkyl.

[148] The compound of paragraph [141], or pharmaceutically acceptablesalt thereof, having the structure of Formula VIb:

wherein a is 0 or 1.

[149] The compound of paragraph [148], or pharmaceutically acceptablesalt thereof, having the structure of Formula VIb-1:

wherein X² is N or CH;

each R³ is independently selected from halogen, cyano, hydroxy,optionally substituted amine, optionally substituted amido, optionallysubstituted C₁-C₆ alkyl, optionally substituted C₁-C₆ heteroalkyl,optionally substituted 3 to 6-membered cycloalkyl, optionallysubstituted 3 to 6-membered cycloalkenyl, optionally substituted 3 to11-membered heterocycloalkyl, optionally substituted 6 to 10-memberedaryl, or optionally substituted 5 to 10-membered heteroaryl; and

n is an integer from 1 to 4.

[150] The compound of paragraph [149], or pharmaceutically acceptablesalt thereof, having the structure of Formula VIb-2:

[151] The compound of paragraph [150], or pharmaceutically acceptablesalt thereof, having the structure of Formula VIb-3:

wherein R⁴ and R⁵ are each independently selected from hydrogen,halogen, cyano, hydroxy, optionally substituted amine, optionallysubstituted amido, optionally substituted C₁-C₆ alkyl, optionallysubstituted C₁-C₆ heteroalkyl, optionally substituted 3 to 6-memberedcycloalkyl, optionally substituted 3 to 6-membered cycloalkenyl,optionally substituted 3 to 11-membered heterocycloalkyl, optionallysubstituted 6 to 10-membered aryl, or optionally substituted 5 to10-membered heteroaryl. In some embodiments, R⁴ and R⁵ are not hydrogen.

[152] The compound of paragraph [151], or pharmaceutically acceptablesalt thereof, having the structure of Formula VIb-4:

[153] The compound of paragraph [152], or pharmaceutically acceptablesalt thereof, having the structure of Formula VIb-5:

wherein X³ is N or CH;

m is 1 or 2;

R⁶, R⁷, R⁸, and R¹¹ are each independently selected from hydrogen,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 6-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl; or

R⁶ and R⁷ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered cycloalkyl or an optionallysubstituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R⁸ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R¹¹ combine with the atoms to which they are attached to form anoptionally substituted 4 to 8-membered heterocycloalkyl.

[154] The compound of paragraph [141], or pharmaceutically acceptablesalt thereof, having the structure of Formula VIIb:

wherein R⁹ is H or C₁-C₆ alkyl; and

a is 0 or 1.

[155] The compound of paragraph [154], or pharmaceutically acceptablesalt thereof, having the structure of Formula VIIb-1:

wherein X² is N or CH;

each R³ is independently selected from halogen, cyano, hydroxy,optionally substituted amine, optionally substituted amido, optionallysubstituted C₁-C₆ alkyl, optionally substituted C₁-C₆ heteroalkyl,optionally substituted 3 to 6-membered cycloalkyl, optionallysubstituted 3 to 6-membered cycloalkenyl, optionally substituted 3 to11-membered heterocycloalkyl, optionally substituted 6 to 10-memberedaryl, or optionally substituted 5 to 10-membered heteroaryl; and

n is an integer from 1 to 4.

[156] The compound of paragraph [155], or pharmaceutically acceptablesalt thereof, having the structure of Formula VIIb-2:

[157] The compound of paragraph [156], or pharmaceutically acceptablesalt thereof, having the structure of Formula VIIb-3:

wherein R⁴ and R⁵ are each independently selected from hydrogen,halogen, cyano, hydroxy, optionally substituted amine, optionallysubstituted amido, optionally substituted C₁-C₆ alkyl, optionallysubstituted C₁-C₆ heteroalkyl, optionally substituted 3 to 6-memberedcycloalkyl, optionally substituted 3 to 6-membered cycloalkenyl,optionally substituted 3 to 11-membered heterocycloalkyl, optionallysubstituted 6 to 10-membered aryl, or optionally substituted 5 to10-membered heteroaryl. In some embodiments, R⁴ and R⁵ are not hydrogen.

[158] The compound of paragraph [157], or pharmaceutically acceptablesalt thereof, having the structure of Formula VIIb-4:

[159] The compound of paragraph [158], or pharmaceutically acceptablesalt thereof, having the structure of Formula VIIb-5:

wherein X³ is N or CH;

m is 1 or 2;

R⁶, R⁷, R⁸, and R¹¹ are each independently selected from hydrogen,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 6-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl; or

R⁶ and R⁷ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered cycloalkyl or an optionallysubstituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R⁸ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R¹¹ combine with the atoms to which they are attached to form anoptionally substituted 4 to 8-membered heterocycloalkyl.

[160] The compound of any one of paragraphs [154]-[159], orpharmaceutically acceptable salt thereof, wherein R⁹ is methyl.

[161] The compound of any one of paragraphs [101]-[108], orpharmaceutically acceptable salt thereof, wherein Y is —NHS(O)— or—NHS(O)NH—.

[162] The compound of paragraph [161], or pharmaceutically acceptablesalt thereof, having the structure of Formula VIIIb:

wherein a is 0 or 1.

[163] The compound of paragraph [162], or pharmaceutically acceptablesalt thereof, having the structure of Formula VIIIb-1:

wherein X² is N or CH;

each R³ is independently selected from halogen, cyano, hydroxy,optionally substituted amine, optionally substituted amido, optionallysubstituted C₁-C₆ alkyl, optionally substituted C₁-C₆ heteroalkyl,optionally substituted 3 to 6-membered cycloalkyl, optionallysubstituted 3 to 6-membered cycloalkenyl, optionally substituted 3 to11-membered heterocycloalkyl, optionally substituted 6 to 10-memberedaryl, or optionally substituted 5 to 10-membered heteroaryl; and

n is an integer from 1 to 4.

[164] The compound of paragraph [163], or pharmaceutically acceptablesalt thereof, having the structure of Formula VIIIb-2:

[165] The compound of paragraph [164], or pharmaceutically acceptablesalt thereof, having the structure of Formula VIIIb-3:

wherein R⁴ and R⁵ are each independently selected from hydrogen,halogen, cyano, hydroxy, optionally substituted amine, optionallysubstituted amido, optionally substituted C₁-C₆ alkyl, optionallysubstituted C₁-C₆ heteroalkyl, optionally substituted 3 to 6-memberedcycloalkyl, optionally substituted 3 to 6-membered cycloalkenyl,optionally substituted 3 to 11-membered heterocycloalkyl, optionallysubstituted 6 to 10-membered aryl, or optionally substituted 5 to10-membered heteroaryl. In some embodiments, R⁴ and R⁵ are not hydrogen.

[166] The compound of paragraph [165], or pharmaceutically acceptablesalt thereof, having the structure of Formula VIIIb-4:

[167] The compound of paragraph [166], or pharmaceutically acceptablesalt thereof, having the structure of Formula VIIIb-5:

wherein X³ is N or CH;

m is 1 or 2;

R⁶, R⁷, R⁸, and R¹¹ are each independently selected from hydrogen,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 6-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl; or

R⁶ and R⁷ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered cycloalkyl or an optionallysubstituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R⁸ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R¹¹ combine with the atoms to which they are attached to form anoptionally substituted 4 to 8-membered heterocycloalkyl.

[168] The compound of paragraph [161], or pharmaceutically acceptablesalt thereof, having the structure of Formula IXb:

wherein a is 0 or 1.

[169], The compound of paragraph [168], or pharmaceutically acceptablesalt thereof, having the structure of Formula IXb-1:

wherein X² is N or CH;

each R³ is independently selected from halogen, cyano, hydroxy,optionally substituted amine, optionally substituted amido, optionallysubstituted C₁-C₆ alkyl, optionally substituted C₁-C₆ heteroalkyl,optionally substituted 3 to 6-membered cycloalkyl, optionallysubstituted 3 to 6-membered cycloalkenyl, optionally substituted 3 to11-membered heterocycloalkyl, optionally substituted 6 to 10-memberedaryl, or optionally substituted 5 to 10-membered heteroaryl; and

n is an integer from 1 to 4.

[170] The compound of paragraph [169], or pharmaceutically acceptablesalt thereof, having the structure of Formula IXb-2:

[171] The compound of paragraph [170], or pharmaceutically acceptablesalt thereof, having the structure of Formula IXb-3:

wherein R⁴ and R⁵ are each independently selected from hydrogen,halogen, cyano, hydroxy, optionally substituted amine, optionallysubstituted amido, optionally substituted C₁-C₆ alkyl, optionallysubstituted C₁-C₆ heteroalkyl, optionally substituted 3 to 6-memberedcycloalkyl, optionally substituted 3 to 6-membered cycloalkenyl,optionally substituted 3 to 11-membered heterocycloalkyl, optionallysubstituted 6 to 10-membered aryl, or optionally substituted 5 to10-membered heteroaryl. In some embodiments, R⁴ and R⁵ are not hydrogen.

[172] The compound of paragraph [171], or pharmaceutically acceptablesalt thereof, having the structure of Formula IXb-4:

[173] The compound of paragraph [172], or pharmaceutically acceptablesalt thereof, having the structure of Formula IXb-5:

wherein X³ is N or CH;

m is 1 or 2;

R⁶, R⁷, R⁸, and R¹¹ are each independently selected from hydrogen,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 6-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl; or

R⁶ and R⁷ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered cycloalkyl or an optionallysubstituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R⁸ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R¹¹ combine with the atoms to which they are attached to form anoptionally substituted 4 to 8-membered heterocycloalkyl.

[174] The compound of paragraph [161], or pharmaceutically acceptablesalt thereof, having the structure of Formula Xb:

wherein R⁹ is H or C₁-C₆ alkyl; and

a is 0 or 1.

[174] The compound of paragraph [174], or pharmaceutically acceptablesalt thereof, having the structure of Formula Xb-1:

wherein X² is N or CH;

each R³ is independently selected from halogen, cyano, hydroxy,optionally substituted amine, optionally substituted amido, optionallysubstituted C₁-C₆ alkyl, optionally substituted C₁-C₆ heteroalkyl,optionally substituted 3 to 6-membered cycloalkyl, optionallysubstituted 3 to 6-membered cycloalkenyl, optionally substituted 3 to11-membered heterocycloalkyl, optionally substituted 6 to 10-memberedaryl, or optionally substituted 5 to 10-membered heteroaryl; and

n is an integer from 1 to 4.

[176] The compound of paragraph [175], or pharmaceutically acceptablesalt thereof, having the structure of Formula Xb-2;

[177] The compound of paragraph [176], or pharmaceutically acceptablesalt thereof, having the structure of Formula Xb-3:

wherein R⁴ and R⁵ are each independently selected from hydrogen,halogen, cyano, hydroxy, optionally substituted amine, optionallysubstituted amido, optionally substituted C₁-C₆ alkyl, optionallysubstituted C₁-C₆ heteroalkyl, optionally substituted 3 to 6-memberedcycloalkyl, optionally substituted 3 to 6-membered cycloalkenyl,optionally substituted 3 to 11-membered heterocycloalkyl, optionallysubstituted 6 to 10-membered aryl, or optionally substituted 5 to10-membered heteroaryl. In some embodiments, R⁴ and R⁵ are not hydrogen.

[178] The compound of paragraph [177], or pharmaceutically acceptablesalt thereof, having the structure of Formula Xb-4:

[179] The compound of paragraph [178], or pharmaceutically acceptablesalt thereof, having the structure of Formula Xb-5:

wherein X³ is N or CH;

m is 1 or 2;

R⁶, R⁷, R⁸, and R¹¹ are each independently selected from hydrogen,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 6-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl; or

R⁶ and R⁷ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered cycloalkyl or an optionallysubstituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R⁸ combine with the atoms to which they are attached to form anoptionally substituted 3 to 8-membered heterocycloalkyl; or

R⁷ and R¹¹ combine with the atoms to which they are attached to form anoptionally substituted 4 to 8-membered heterocycloalkyl.

[180] The compound of any one of paragraphs [174]-[179], orpharmaceutically acceptable salt thereof, wherein R⁹ is methyl.

[181] The compound of any one of paragraphs [110]-[140], [142]-[160], or[162]-[180], or pharmaceutically acceptable salt thereof, wherein a is0.

[182] The compound of any one of paragraphs [110]-[140], [142]-[160], or[162]-[180], or pharmaceutically acceptable salt thereof, wherein a is1.

[183] The compound of any one of paragraphs [101]-[182], orpharmaceutically acceptable salt thereof, wherein R² is optionallysubstituted C₁-C₆ alkyl.

[184] The compound of paragraph [183], or pharmaceutically acceptablesalt thereof, wherein R² is selected from —CH₂CH₃ or—CH₂CF₃.

[185] The compound of any one of paragraphs [101]-[184], orpharmaceutically acceptable salt thereof, wherein R¹⁰ is hydrogen.

[186] The compound of any one of paragraphs [101]-[184], orpharmaceutically acceptable salt thereof, wherein R¹⁰ is hydroxy,optionally substituted C₁-C₆ alkoxy, optionally substituted C₁-C₃ alkyl,optionally substituted C₁-C₆ heteroalkyl, or optionally substituted 3 to7-membered heterocycloalkyl.

[187] The compound of paragraph [186], or pharmaceutically acceptablesalt thereof, wherein R¹⁰ is optionally substituted ethoxy.

[188] The compound of paragraph [186], or pharmaceutically acceptablesalt thereof, wherein R¹⁰ is selected from the following, or astereoisomer thereof:

[189] The compound of any one of paragraphs [101]-[188], orpharmaceutically acceptable salt thereof, wherein W is C₁-C₄ alkyl.

[190] The compound of any one of paragraphs [101]-[188], orpharmaceutically acceptable salt thereof, wherein W is optionallysubstituted cyclopropyl, optionally substituted cyclobutyl, optionallysubstituted cyclopentyl, or optionally substituted cyclohexyl,optionally substituted piperidine, optionally substituted piperazine,optionally substituted pyridine, or optionally substituted phenyl.

[191] The compound of any one of paragraphs [101]-[188], orpharmaceutically acceptable salt thereof, wherein W is optionallysubstituted 3 to 10-membered heterocycloalkyl, optionally substituted 3to 10-membered cycloalkyl, optionally substituted 6 to 10-membered aryl,or optionally substituted 5 to 10-membered heteroaryl.

[192] The compound of any one of paragraphs [101]-[188], orpharmaceutically acceptable salt thereof, wherein W is optionallysubstituted 3 to 10-membered heterocycloalkyl.

[193] The compound of paragraph [192], or pharmaceutically acceptablesalt thereof, wherein W is selected from the following, or astereoisomer thereof:

[194] The compound of any one of paragraphs [101]-[188], orpharmaceutically acceptable salt thereof, wherein W is optionallysubstituted 3 to 10-membered cycloalkyl.

[195] The compound of paragraph [194], or pharmaceutically acceptablesalt thereof, wherein W is selected from the following, or astereoisomer thereof:

[196] The compound of any one of paragraphs [101]-[188], orpharmaceutically acceptable salt thereof, wherein W is optionallysubstituted 5 to 10-membered heteroaryl.

[197] The compound of paragraph [196], or pharmaceutically acceptablesalt thereof, wherein W is selected from the following, or astereoisomer thereof:

[198] The compound of any one of paragraphs [101]-[188], orpharmaceutically acceptable salt thereof, wherein W is optionallysubstituted 6 to 10-membered aryl.

[199] The compound of paragraph [198], or pharmaceutically acceptablesalt thereof, wherein W is optionally substituted phenyl.

[200] The compound of any one of paragraphs [101]-[188], orpharmaceutically acceptable salt thereof, wherein W is optionallysubstituted C₁-C₃ heteroalkyl.

[201] The compound of paragraph [200], or pharmaceutically acceptablesalt thereof, wherein W is selected from the following, or astereoisomer thereof:

[202] The compound of paragraph [189], or pharmaceutically acceptablesalt thereof, wherein W is selected from the following:

[203] The compound of any one of paragraphs [101]-[188], orpharmaceutically acceptable salt thereof, wherein W is —R¹⁴C(═O)R¹⁵where R¹⁴ is 3 to 10-membered cycloalkylene and R¹⁵ is selected fromoptionally substituted 3 to 10-membered cycloalkyl, optionallysubstituted 6 to 10-membered aryl, or optionally substituted 5 to10-membered heteroaryl.

[204] The compound of paragraph [203], or pharmaceutically acceptablesalt thereof, wherein W is selected from the following, or astereoisomer thereof:

[205] A compound, or a pharmaceutically acceptable salt thereof, ofTable 1a or Table 1b.

[206] A pharmaceutical composition comprising a compound, or apharmaceutically acceptable salt thereof, of any one of paragraphs[1]-[205] and a pharmaceutically acceptable excipient.

[207] A method of treating cancer in a subject in need thereof, themethod comprising administering to the subject a therapeuticallyeffective amount of a compound, or a pharmaceutically acceptable saltthereof, of any one of paragraphs [1]-[205] or a pharmaceuticalcomposition of paragraph [206].

[208] The method of paragraph [207], wherein the cancer comprises a Rasmutation.

[209] The method of paragraph [208], wherein the Ras mutation is atposition 12, 13 or 61. In some embodiments, the Ras mutation is atposition 12.

[210] The method of paragraph [209], wherein the Ras mutation is at aposition selected from the group consisting of G12C, G12D, G12V, G12R,G13C, G13D, and Q61K, or a combination thereof.

[211] The method of paragraph [210], wherein the Ras mutation is at aposition selected from the group consisting of G12D, G12V and G12R, or acombination thereof.

[212] The method of paragraph [211], wherein the Ras mutation is at aposition selected from the group consisting of G12D and G12V, or acombination thereof.

[213] The method of any one of paragraphs [207]-[212], wherein thecancer is pancreatic cancer.

[214] The method of any one of paragraphs [207]-[212], wherein thecancer is lung cancer.

[215] The method of any one of paragraphs [207]-[212], wherein thecancer is colorectal cancer.

[216] A method of treating a Ras protein-related disorder in a subjectin need thereof, the method comprising administering to the subject atherapeutically effective amount of a compound, or a pharmaceuticallyacceptable salt thereof, of any one of paragraphs [1]-[205] or apharmaceutical composition of paragraph [206].

[217] A method of inhibiting a Ras protein in a cell, the methodcomprising contacting the cell with an effective amount of a compound,or a pharmaceutically acceptable salt thereof, of any one of paragraphs[1]-[205] or a pharmaceutical composition of paragraph [206].

The method of paragraph [217], wherein more than one Ras protein isinhibited in the cell.

The method of [217] or [218], wherein the cell is a cancer cell.

The method of paragraph [219], wherein the cancer cell is a pancreaticcancer cell.

The method of paragraph [219], wherein the cancer cell is a lung cancercell.

The method of paragraph [219], wherein the cancer cell is a colorectalcancer cell.

The method of any one of paragraphs [207]-[222], wherein the Ras proteinis KRAS.

The method or use of any one of paragraphs [207]-[223], wherein themethod further comprises administering an additional anticancer therapy.

The method of paragraph [224], wherein the additional anticancer therapyis an EGFR inhibitor, a second Ras inhibitor, a SHP2 inhibitor, a SOS1inhibitor, a Raf inhibitor, a MEK inhibitor, an ERK inhibitor, a PI3Kinhibitor, a PTEN inhibitor, an AKT inhibitor, an mTORC1 inhibitor, aBRAF inhibitor, a PD-L1 inhibitor, a PD-1 inhibitor, a CDK4/6 inhibitor,a HER2 inhibitor, or a combination thereof.

The method of paragraph [224] or [225], wherein the additionalanticancer therapy is a SHP2 inhibitor.

The method of paragraph [224] or [225], wherein the additionalanticancer therapy comprises a SHP2 inhibitor and a PD-L1 inhibitor.

The method of paragraph [224] or [225], wherein the the additionaltherapy comprises a second Ras inhibitor and a PD-L1 inhibitor.

The method of paragraph [225] or [228], wherein the second Ras inhibitoris a KRAS^(G12C) inhibitor.

The method of paragraph [228] or [229], wherein the second Ras inhibitoris a KRAS^(G12C)(ON) inhibitor.

The method of paragraph [228] or [229], wherein the second Ras inhibitoris a KRAS^(G12C)(OFF) inhibitor.

EXAMPLES

The disclosure is further illustrated by the following examples andsynthesis examples, which are not to be construed as limiting thisdisclosure in scope or spirit to the specific procedures hereindescribed. It is to be understood that the examples are provided toillustrate certain embodiments and that no limitation to the scope ofthe disclosure is intended thereby. It is to be further understood thatresort may be had to various other embodiments, modifications, andequivalents thereof which may suggest themselves to those skilled in theart without departing from the spirit of the present disclosure or scopeof the appended claims.

Chemical Syntheses

Definitions used in the following examples and elsewhere herein are:

-   -   B₂pin₂ Bis(pinacolato)diboron    -   BINAP 2,2′-Bis(diphenylphosphino)-1,1′-binaphthyl    -   CH₂Cl₂, DCM Methylene chloride, Dichloromethane    -   CH₃CN, MeCN Acetonitrile    -   CuI Copper (I) iodide    -   DIPEA, DIEA Diisopropylethyl amine    -   DMF N,N-Dimethylformamide    -   EA Ethyl acetate    -   EDCl N-Ethyl-N′-carbodiimide hydrochloride    -   EtOAc Ethyl acetate    -   h hour    -   H₂O Water    -   HCl Hydrochloric acid    -   HOBt Hydroxybenzotriazole    -   K₃PO₄ Potassium phosphate (tribasic)    -   MeOH Methanol    -   Na₂SO₄ Sodium sulfate    -   NMM N-methylmorpholine    -   NMP N-methyl pyrrolidone    -   Pd(dppf)Cl₂        [1,1′-Bis(diphenylphosphino)ferrocene]dichloropalladium(II)    -   PE Petroleum ether    -   rt Room temperature    -   TFA Trifluoroacetic acid

Instrumentation

Mass spectrometry data collection took place with a Shimadzu LCMS-2020,an Agilent 1260LC-6120/6125MSD, a Shimadzu LCMS-2010EV, or a WatersAcquity UPLC, with either a QDa detector or SQ Detector 2. Samples wereinjected in their liquid phase onto a C-18 reverse phase. The compoundswere eluted from the column using an acetonitrile gradient and fed intothe mass analyzer. Initial data analysis took place with either AgilentChemStation, Shimadzu LabSolutions, or Waters MassLynx. NMR data wascollected with either a Bruker AVANCE III HD 400 MHz, a Bruker Ascend500 MHz instrument, or a Varian 400 MHz, and the raw data was analyzedwith either TopSpin or Mestrelab Mnova.

Synthesis of Intermediates Intermediate 1. Synthesis of3-(5-bromo-1-ethyl-2-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]indol-3-yl)-2,2-dimethylpropan-1-ol

Step 1

To a mixture of 3-(tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropanoylchloride (65 g, 137 mmol, crude) in DCM (120 mL) at 0° C. under anatmosphere of N₂ was added 1M SnCl₄ in DCM (137 mL, 137 mmol) slowly.The mixture was stirred at 0° C. for 30 min, then a solution of5-bromo-1H-indole (26.8 g, 137 mmol) in DCM (40 mL) was added dropwise.The mixture was stirred at 0° C. for 45 min, then diluted with EtOAc(300 mL), washed with brine (400 mL), dried over Na₂SO₄ and filtered.The filtrate was concentrated under reduced pressure and the residue waspurified by silica gel column chromatography to give1-(5-bromo-1H-indol-3-yl)-3-(tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropan-1-one(55 g, 75% yield). LCMS (ESI): m/z [M+Na] calc'd for C₂₉H₃₂BrNO₂SiNa556.1; found 556.3.

Step 2

To a mixture of1-(5-bromo-1H-indol-3-yl)-3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropan-1-one(50 g, 93.6 mmol) in THF (100 mL) at 0° C. under an atmosphere of N₂ wasadded LiBH₄ (6.1 g, 281 mmol). The mixture was heated to 60° C. andstirred for 20 h, then MeOH (10 mL) and EtOAc (100 mL) were added andthe mixture washed with brine (50 mL), dried over Na₂SO₄, filtered andthe filtrate concentrated under reduced pressure. The residue wasdiluted with DCM (50 mL), cooled to 10° C. and diludine (9.5 g, 37.4mmol) and TsOH. H₂O (890 mg, 4.7 mmol) added. The mixture was stirred at10° C. for 2 h, filtered, the filtrate concentrated under reducedpressure and the residue was purified by silica gel columnchromatography to give1-(5-bromo-1H-indol-3-yl)-3-(tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropan-1-one(41 g, 84% yield). LCMS (ESI): m/z [M+H] calc'd for C₂₉H₃₄BrNOSi 519.2;found 520.1; ¹H NMR (400 MHz, CDCl₃) δ 7.96 (s, 1H), 7.75-7.68 (m, 5H),7.46-7.35 (m, 6H), 7.23-7.19 (m, 2H), 6.87 (d, J=2.1 Hz, 1H), 3.40 (s,2H), 2.72 (s, 2H), 1.14 (s, 9H), 0.89 (s, 6H).

Step 3

To a mixture of1-(5-bromo-1H-indol-3-yl)-3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropan-1-one(1.5 g, 2.9 mmol) and I₂ (731 mg, 2.9 mmol) in THF (15 mL) at rt wasadded AgOTf (888 mg, 3.5 mmol). The mixture was stirred at rt for 2 h,then diluted with EtOAc (200 mL) and washed with saturated Na₂S₂O₃ (100mL), dried over anhydrous Na₂SO₄ and filtered. The filtrate wasconcentrated under reduced pressure and the residue was purified bysilica gel column chromatography to give5-bromo-3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-2-iodo-1H-indole(900 mg, 72% yield) as a solid. ¹H NMR (400 MHz, DMSO-d₆) δ 11.70 (s,1H), 7.68 (d, J=1.3 Hz, 1H), 7.64-7.62 (m, 4H), 7.46-7.43 (m, 6H),7.24-7.22 (d, 1H), 7.14-7.12 (dd, J=8.6, 1.6 Hz, 1H), 3.48 (s, 2H), 2.63(s, 2H), 1.08 (s, 9H), 0.88 (s, 6H).

Step 4

To a stirred mixture of HCOOH (66.3 g, 1.44 mol) in TEA (728 g, 7.2 mol)at 0° C. under an atmosphere of Ar was added(4S,5S)-2-chloro-2-methyl-1-(4-methylbenzenesulfonyl)-4,5-diphenyl-1,3-diaza-2-ruthenacyclopentanecymene (3.9 g, 6.0 mmol) portion-wise. The mixture was heated to 40° C.and stirred for 15 min, then cooled to rt and1-(3-bromopyridin-2-yl)ethanone (120 g, 600 mmol) added in portions. Themixture was heated to 40° C. and stirred for an additional 2 h, then thesolvent was concentrated under reduced pressure. Brine (2 L) was addedto the residue, the mixture was extracted with EtOAc (4×700 mL), driedover anhydrous Na₂SO₄ and filtered. The filtrate was concentrated underreduced pressure and the residue was purified by silica gel columnchromatography to give (1S)-1-(3-bromopyridin-2-yl)ethanol (100 g, 74%yield) as an oil. LCMS (ESI): m/z [M+H] calc'd for C₇H₈BrNO 201.1; found201.9.

Step 5

To a stirred mixture of (1S)-1-(3-bromopyridin-2-yl)ethanol (100 g, 495mmol) in DMF (1 L) at 0° C. was added NaH, 60% dispersion in oil (14.25g, 594 mmol) in portions. The mixture was stirred at 0° C. for 1 h. Mel(140.5 g, 990 mmol) was added dropwise at 0° C. and the mixture wasallowed to warm to rt and stirred for 2 h. The mixture was cooled to 0°C. and saturated NH₄Cl (5 L) was added. The mixture was extracted withEtOAc (3×1.5 L), dried over anhydrous Na₂SO₄ and filtered. The filtratewas concentrated under reduced pressure and the residue was purified bysilica gel column chromatography to give3-bromo-2-[(1S)-1-methoxyethyl]pyridine (90 g, 75% yield) as an oil.LCMS (ESI): m/z [M+H] calc'd for C₈H₁₀BrNO 215.0; found 215.9.

Step 6

To a stirred mixture of 3-bromo-2-[(1S)-1-methoxyethyl]pyridine (90 g,417 mmol) and Pd(dppf)Cl₂ (30.5 g, 41.7 mmol) in toluene (900 mL) at rtunder an atmosphere of Ar was added bis(pinacolato)diboron (127 g, 500mmol) and KOAc (81.8 g, 833 mmol) in portions. The mixture was heated to100° C. and stirred for 3 h. The filtrate was concentrated under reducedpressure and the residue was purified by Al₂O₃ column chromatography togive2-[(1S)-1-methoxyethyl]-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine(100 g, 63% yield) as a semi-solid. LCMS (ESI): m/z [M+H] calc'd forC₁₄H₂₂BNO₃ 263.2; found 264.1.

Step 7

To a stirred mixture of5-bromo-3-[3-[(tert-butyldiphenylsilyl)oxy]-2,2-dimethylpropyl]-2-iodo-1H-indole(140 g, 217 mmol) and2-[(1S)-1-methoxyethyl]-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine(100 g, 380 mmol) in 1,4-dioxane (1.4 L) at rt under an atmosphere of Arwas added K₂CO₃ (74.8 g, 541 mmol), Pd(dppf)Cl₂ (15.9 g, 21.7 mmol) andH₂O (280 mL) in portions. The mixture was heated to 85° C. and stirredfor 4 h, then cooled, H₂O (5 L) added and the mixture extracted withEtOAc (3×2 L). The combined organic layers were washed with brine (2×1L), dried over anhydrous Na₂SO₄ and filtered. The filtrate wasconcentrated under reduced pressure and the residue was purified bysilica gel column chromatography to give5-bromo-3-[3-[(tert-butyldiphenylsilyl)oxy]-2,2-dimethylpropyl]-2-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]-1H-indole(71 g, 45% yield) as a solid. LCMS (ESI): m/z [M+H] calc'd forC₃₇H₄₃BrN₂O₂Si, 654.2; found 655.1.

Step 8

To a stirred mixture of5-bromo-3-[3-[(tert-butyldiphenylsilyl)oxy]-2,2-dimethylpropyl]-2-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]-1H-indole(71 g, 108 mmol) in DMF (0.8 L) at 0° C. under an atmosphere of N₂ wasadded Cs₂CO₃ (70.6 g, 217 mmol) and EtI (33.8 g, 217 mmol) in portions.The mixture was warmed to rt and stirred for 16 h then H₂O (4 L) addedand the mixture extracted with EtOAc (3×1.5 L). The combined organiclayers were washed with brine (2×1 L), dried over anhydrous Na₂SO₄ andfiltered. The filtrate was concentrated under reduced pressure and theresidue was purified by silica gel column chromatography to give5-bromo-3-[3-[(tert-butyldiphenylsilyl)oxy]-2,2-dimethylpropyl]-1-ethyl-2-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]indole(66 g, 80% yield) as an oil. LCMS (ESI): m/z [M+H] calc'd forC₃₉H₄₇BrN₂O₂Si, 682.3; found 683.3.

Step 9

To a stirred mixture of TBAF (172.6 g, 660 mmol) in THF (660 mL) at rtunder an atmosphere of N₂ was added5-bromo-3-[3-[(tert-butyldiphenylsilyl)oxy]-2,2-dimethylpropyl]-1-ethyl-2-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]indole(66 g, 97 mmol) in portions. The mixture was heated to 50° C. andstirred for 16 h, cooled, diluted with H₂O (5 L) and extracted withEtOAc (3×1.5 L). The combined organic layers were washed with brine (2×1L), dried over anhydrous Na₂SO₄ and filtered. After filtration, thefiltrate was concentrated under reduced pressure. The filtrate wasconcentrated under reduced pressure and the residue was purified bysilica gel column chromatography to give two atropisomers of3-(5-bromo-1-ethyl-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-indol-3-yl)-2,2-dimethylpropan-1-ol(as single atropisomers) both as solids. (Combined 30 g, 62% yield) bothas a solid. LCMS (ESI): m/z [M+H] calc'd for C₂₃H₂₉BrN₂O₂ 444.1; found445.1.

Intermediate 1. Alternative Synthesis Through Fisher Indole Route

Step 1

To a mixture of i-PrMgCl (2M in in THF, 0.5 L) at −10° C. under anatmosphere of N₂ was added n-BuLi, 2.5 M in hexane (333 mL, 833 mmol)dropwise over 15 min. The mixture was stirred for 30 min at −10° C. then3-bromo-2-[(1S)-1-methoxyethyl]pyridine (180 g, 833 mmol) in THF (0.5 L)added dropwise over 30 min at −10° C. The resulting mixture was warmedto −5° C. and stirred for 1 h, then 3,3-dimethyloxane-2,6-dione (118 g,833 mmol) in THF (1.2 L) was added dropwise over 30 min at −5° C. Themixture was warmed to 0° C. and stirred for 1.5 h, then quenched withthe addition of pre-cooled 4M HCl in 1,4-dioxane (0.6 L) at 0° C. toadjust pH ˜5. The mixture was diluted with ice-water (3 L) and extractedwith EtOAc (3×2.5 L). The combined organic layers were dried overanhydrous Na₂SO₄, filtered, the filtrate was concentrated under reducedpressure and the residue was purified by silica gel columnchromatography to give5-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]-2,2-dimethyl-5-oxopentanoic acid(87 g, 34% yield) as a solid. LCMS (ESI): m/z [M+H] calc'd for C₁₅H₂₁NO₄279.2; found 280.1.

Step 2

To a mixture of5-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]-2,2-dimethyl-5-oxopentanoic acid(78 g, 279 mmol) in EtOH (0.78 L) at rt under an atmosphere of N₂ wasadded (4-bromophenyl)hydrazine HCl salt (68.7 g, 307 mmol) in portions.The mixture was heated to 85° C. and stirred for 2 h, cooled to rt, then4M HCl in 1,4-dioxane (69.8 mL, 279 mmol) added dropwise. The mixturewas heated to 85° C. and stirred for an additional 3 h, thenconcentrated under reduced pressure and the residue was dissolved in TFA(0.78 L). The mixture was heated to 60° C. and stirred for 1.5,concentrated under reduced pressure and the residue adjusted to pH ˜5with saturated NaHCO₃, then extracted with EtOAc (3×1.5 L). The combinedorganic layers were dried over anhydrous Na₂SO₄, filtered, the filtrateconcentrated under reduced pressure and the residue was purified bysilica gel column chromatography to give3-(5-bromo-2-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]-1H-indol-3-yl)-2,2-dimethylpropanoicacid and ethyl(S)-3-(5-bromo-2-(2-(1-methoxyethyl)pyridin-3-yl)-1H-indol-3-yl)-2,2-dimethylpropanoate(78 g, crude). LCMS (ESI): m/z [M+H] calc'd for C₂₁H₂₃BrN₂O₃ 430.1 andC₂₃H₂₇BrN₂O₃ 458.1; found 431.1 and 459.1.

Step 3

To a mixture of3-(5-bromo-2-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]-1H-indol-3-yl)-2,2-dimethylpropanoicacid and ethyl(S)-3-(5-bromo-2-(2-(1-methoxyethyl)pyridin-3-yl)-1H-indol-3-yl)-2,2-dimethylpropanoate(198 g, 459 mmol) in DMF (1.8 L) at 0° C. under an atmosphere of N₂ wasadded Cs₂CO₃ (449 g, 1.38 mol) in portions. EtI (215 g, 1.38 mmol) inDMF (200 mL) was then added dropwise at 0° C. The mixture was warmed tort and stirred for 4 h then diluted with brine (5 L) and extracted withEtOAc (3×2.5 L). The combined organic layers were washed with brine(2×1.5 L), dried over anhydrous Na₂SO₄ and filtered. The filtrate wasconcentrated under reduced pressure and the residue was purified bysilica gel column chromatography to give ethyl3-(5-bromo-1-ethyl-2-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]indol-3-yl)-2,2-dimethylpropanoate(160 g, 57% yield) as a solid. LCMS (ESI): m/z [M+H] calc'd forC₂₅H₃₁BrN₂CO₃ 486.2; found 487.2.

Step 4

To a mixture of ethyl3-(5-bromo-1-ethyl-2-[2-[(1S)-1-methoxyethyl]pyridin-3-yl]indol-3-yl)-2,2-dimethylpropanoate(160 g, 328 mmol) in THF (1.6 L) at 0° C. under an atmosphere of N₂ wasadded LiBH₄ (28.6 g, 1.3 mol). The mixture was heated to 60° C. for 16h, cooled, and quenched with pre-cooled (0° C.) aqueous NH₄Cl (5 L). Themixture was extracted with EtOAc (3×2 L) and the combined organic layerswere washed with brine (2×1 L), dried over anhydrous Na₂SO₄ andfiltered. The filtrate was concentrated under reduced pressure and theresidue was purified by silica gel column chromatography to give to twoatropisomers of3-(5-bromo-1-ethyl-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-indol-3-yl)-2,2-dimethylpropan-1-ol(as single atropisomers) (60 g, 38% yield) and (40 g, 26% yield) both assolids. LCMS (ESI): m/z [M+H] calc'd for C₂₃H₂₉BrN₂O₂ 444.1; found445.2.

Intermediate 2. Synthesis of tert-butyl((6³S,4S,Z)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate

Step 1

To a solution of methyl(2S)-3-(4-bromo-1,3-thiazol-2-yl)-2-[(tert-butoxycarbonyl)amino]propanoate(110 g, 301.2 mmol) in THF (500 mL) and H₂O (200 mL) at room temperaturewas added LiOH (21.64 g, 903.6 mmol). The solution was stirred for 1 hand was then concentrated under reduced pressure. The residue wasadjusted to pH 6 with 1 M HCl and then extracted with DCM (3×500 mL).The combined organic layers were, dried over Na₂SO₄, filtered, andconcentrated under reduced pressure to give(S)-3-(4-bromothiazol-2-yl)-2-((tert-butoxycarbonyl)amino)propanoic acid(108 g, crude). LCMS (ESI): m/z [M+H] calc'd for C₁₁H₁₆BrN₂O₄S. 351.0;found 351.0.

Step 2

To a solution of(S)-3-(4-bromothiazol-2-yl)-2-((tert-butoxycarbonyl)amino)propanoic acid(70 g, 199.3 mmol) in DCM (500 mL) at 0° C. was added methyl(3S)-1,2-diazinane-3-carboxylate bis(trifluoroacetic acid) salt (111.28g, 298.96 mmol), NMM (219.12 mL. 1993.0 mmol), EDCl (76.41 g, 398.6mmol) and HOBt (5.39 g, 39.89 mmol). The solution was warmed to roomtemperature and stirred for 1 h. The reaction was then quenched with H₂O(500 mL) and was extracted with EtOAc (3×500 mL). The combined organiclayers were dried over Na₂SO₄, filtered, and concentrated under reducedpressured. The residue was purified by silica gel column chromatographyto give methyl(S)-1-((S)-3-(4-bromothiazol-2-yl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylate(88.1 g, 93% yield). LCMS (ESI): m/z [M+H] calc'd for C₁₇H₂₆BrN₄O₅S.477.1; found 477.1.

Step 3

To a solution of3-(5-bromo-1-ethyl-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-indol-3-yl)-2,2-dimethylpropan-1-ol(60 g, 134.7 mmol) in toluene (500 mL) at room temperature was addedbis(pinacolato)diboron (51.31 g, 202.1 mmol), Pd(dppf)Cl₂ (9.86 g, 13.4mmol), and KOAc (26.44 g, 269 mmol). The reaction mixture was thenheated to 90° C. and stirred for 2 h. The reaction solution was thencooled to room temperature and concentrated under reduced pressure. Theresidue was purified by silica gel column chromatography to give(S)-3-(1-ethyl-2-(2-(1-methoxyethyl)pyridin-3-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)-2,2-dimethylpropan-1-ol(60.6 g, 94% yield). LCMS (ESI): m/z [M+H] calc'd for C₂₉H₄₂BN₂O₄493.32; found 493.3.

Step 4

To a solution of(S)-3-(1-ethyl-2-(2-(1-methoxyethyl)pyridin-3-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)-2,2-dimethylpropan-1-ol(30 g, 60.9 mmol) in toluene (600 mL), dioxane (200 mL), and H₂O (200mL) at room temperature was added methyl(S)-1-((S)-3-(4-bromothiazol-2-yl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylate(43.62 g, 91.4 mmol), K₃PO₄ (32.23 g, 152.3 mmol) and Pd(dppf)Cl₂ (8.91g, 12.18 mmol). The resulting solution was heated to 70° C. and stirredovernight. The reaction mixture was then cooled to room temperature andwas quenched with H₂O (200 mL). The mixture was extracted with EtOAc andthe combined organic layers were dried over Na₂SO₄, filtered, andconcentrated under reduced pressure. The residue was purified by silicagel column chromatography to give methyl(S)-1-((S)-2-((tert-butoxycarbonyl)amino)-3-(4-(1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-indol-5-yl)thiazol-2-yl)propanoyl)hexahydropyridazine-3-carboxylate(39.7 g, 85% yield). LCMS (ESI): m/z [M+H] calc'd for C₄₀H₅₅N₆O₇S.763.4; found 763.3.

Step 5

To a solution of methyl(S)-1-((S)-2-((tert-butoxycarbonyl)amino)-3-(4-(1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-indol-5-yl)thiazol-2-yl)propanoyl)hexahydropyridazine-3-carboxylate(39.7 g, 52.0 mmol) in THF (400 mL) and H₂O (100 mL) at room temperaturewas added LiOH.H₂O (3.74 g, 156.2 mmol). The mixture was stirred for 1.5h and was then concentrated under reduced pressure. The residue wasacidified to pH 6 with 1 M HCl and extracted with DCM (3×1000 mL). Thecombined organic layers were dried over Na₂SO₄, filtered, andconcentrated under reduced pressure to give(S)-1-((S)-2-((tert-butoxycarbonyl)amino)-3-(4-(1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-indol-5-yl)thiazol-2-yl)propanoyl)hexahydropyridazine-3-carboxylicacid (37.9 g, crude). LCMS (ESI): m/z [M+H] calc'd for C₃₉H₅₃N₆O₇S.749.4; found 749.4.

Step 6

To a solution of(S)-1-((S)-2-((tert-butoxycarbonyl)amino)-3-(4-(1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1H-indol-5-yl)thiazol-2-yl)propanoyl)hexahydropyridazine-3-carboxylicacid (37.9 g, 50.6 mmol), HOBt (34.19 g, 253.0 mmol) and DIPEA (264.4mL, 1518 mmol) in DCM (4 L) at 0° C. was added EDCl (271.63 g, 1416.9mmol). The resulting mixture was warmed to room temperature and stirredovernight. The reaction mixture was then quenched with H₂O and washedwith 1 M HCl (4×1 L). The organic layer was separated and concentratedunder reduced pressure. The residue was purified by silica gel columnchromatography to give tert-butyl((6³S,4S,Z)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbarnate(30 g, 81% yield). LCMS (ESI): m/z [M+H] calc'd for C₃₉H₅₁N₆O₆S. 731.4;found 731.3.

Step 7

To a solution of tert-butyl((6³S,4S,Z)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(6 g, 8.21 mmol) in DCM (60 mL) at 0° C. was added TFA (30 mL). Themixture was stirred for 1 h and was then concentrated under reducedpressure to give(6³S,4S,Z)-4-amino-1′-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-5,7-dione(7.0 g, crude). LCMS (ESI): m/z [M+H] calc'd for C₃₄H₄₂N₆O₄S. 631.3;found: 630.3.

Intermediate 3. Synthesis of tert-butyl((6³S,4S,Z)-10,10-dimethyl-5,7-dioxo-1²-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate

Step 1

To a stirred solution of5-bromo-3-[3-[(tert-butyldiphenylsilyl)oxy]-2,2-dimethylpropyl]-1H-indole(100 g, 192.0 mmol) in THF (1000 mL) were added TBAF (261.17 g, 998.8mmol) in portion at room temperature. The resulting mixture was stirredfor 16 h at 50° C. The resulting mixture was concentrated under reducedpressure. The resulting mixture was extracted with EtOAc (2 L). Thecombined organic layers were washed with water (6 L), dried overanhydrous Na₂SO₄. After filtration, the filtrate was concentrated underreduced pressure. The residue was purified by silica gel columnchromatography, eluted with PE/EtOAc (3:1) to afford3-(5-bromo-1H-indol-3-yl)-2,2-dimethylpropan-1-ol (54 g, 96.63%). LCMS(ESI): m/z [M+H] calc'd for C₁₃H₁₆BrNO 281.0; found 282.0.

Step 2

To a stirred solution of3-(5-bromo-1H-indol-3-yl)-2,2-dimethylpropan-1-ol (54 g, 191.3 mmol) inDCM (300 mL) were added TEA (58.09 g, 574.1 mmol) and Ac₂O (18.95 g,185.6 mmol) and DMAP (1.17 g, 9.5 mmol) dropwise at 0° C. The resultingmixture was washed with water (3×500 mL), dried over anhydrous Na₂SO₄.After filtration, the filtrate was concentrated under reduced pressure.This resulted in 3-(5-bromo-1H-indol-3-yl)-2,2-dimethylpropyl acetate(54 g, 80.6%) as a yellow solid. The crude product was used in the nextstep directly without further purification. LCMS (ESI): m/z [M+H] calc'dfor C₁₅H₁₈BrNO₂ 323.0; found 324.0.

Step 3

To a stirred solution of 3-(5-bromo-1H-indol-3-yl)-2,2-dimethylpropylacetate (54 g, 166.5 mmol) in toluene (600 mL) were added KOAc (40.87 g,416.3 mmol) and B₂pin₂ (105.76 g, 416.3 mmol) and Pd(dppf)Cl₂ (12.19 g,16.6 mmol) in portions at room temperature under argon atmosphere. Theresulting mixture was stirred for 3 h at 90° C. under argon atmosphere.The resulting mixture was concentrated under reduced pressure. Theresulting mixture was extracted with EtOAc (1 L). The combined organiclayers were washed with water (3×1 L), dried over anhydrous Na₂SO₄.After filtration, the filtrate was concentrated under reduced pressure.The residue was purified by silica gel column chromatography, elutedwith PE/EtOAc (3:1) to afford2,2-dimethyl-3-[5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl]propylacetate borane (55 g, 76.57%) as a yellow solid. LCMS (ESI): m/z [M+H]calc'd for C₂₁H₃₀BNO₄ 371.2; found 372.2.

Step 4

To a stirred solution of2,2-dimethyl-3-[5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-S-yl]propylacetate (54 g, 145.443 mmol) and methyl(2S)-3-(4-bromo-1,3-thiazol-2-yl)-2-[(tert-butoxycarbonyl)amino]propanoate(79.68 g, 218.1 mmol) and K₃PO₄(77.18 g, 363.6 mmol) in toluene (330 mL)and dioxane (110 mL) and H₂O (110 mL) were added Pd(dppf)Cl₂ (10.64 g,14.5 mmol) in portions at room temperature under argon atmosphere. Theresulting mixture was stirred for 36 h at 70° C. under argon atmosphere.The resulting mixture was concentrated under vacuum. The resultingmixture was extracted with EtOAc (3 L). The combined organic layers werewashed with water (3×2 L), dried over anhydrous Na₂SO₄. Afterfiltration, the filtrate was concentrated under reduced pressure. Theresidue was purified by silica gel column chromatography, eluted withPE/EtOAc (1:1) to afford methyl(2S)-3-(4-[3-[3-(acetyloxy)-2,2-dimethylpropyl]-1H-indol-5-yl]-1,3-thiazol-2-yl)-2-[(tert-butoxycarbonyl)amino]propanoate(54 g, 60.78%) as a yellow oil. LCMS (ESI): m/z [M+H] calc'd forC₂₇H₃₅N₃O₆S. 529.2; found 530.2.

Step 5

To a stirred solution of methyl(2S)-3-(4-[3-[3-(acetyloxy)-2,2-dimethylpropyl]-1H-indol-5-yl]-1,3-thiazol-2-yl)-2-[(tert-butoxycarbonyl)amino]propanoate(54 g, 101.954 mmol) in THF (450 mL) were added NaHCO₃ (10.28 g, 122.3mmol) and AgOTf (31.44 g, 122.3 mmol) dropwise at 0° C. To the stirredsolution was added I₂ (23.29 g, 91.6 mmol) in THF (100 mL) dropwise at0° C. The resulting mixture was stirred for 15 min at 0° C. The reactionwas quenched with sat. Na₂S₂O₃ (aq.) at 0° C. The resulting mixture wasextracted with EtOAc (1 L). The combined organic layers were washed withwater (3 L), dried over anhydrous Na₂SO₄. After filtration, the filtratewas concentrated under reduced pressure. The residue was purified bysilica gel column chromatography, eluted with PE/EtOAc (5:1) to affordmethyl(2S)-3-(4-[3-[3-(acetyloxy)-2,2-dimethylpropyl]-2-iodo-1H-indol-5-yl]-1,3-thiazol-2-yl)-2-[(tert-butoxycarbonyl)mino]propanoate (40 g, 53.80%) as a yellow solid. LCMS (ESI): m/z [M+H]calc'd for C₂₇H₃₄IN₃O₆S. 655.1; found 656.1.

Step 6

To a stirred solution of methyl(2S)-3-(4-[3-[3-(acetyloxy)-2,2-dimethylpropyl]-2-iodo-1H-indol-5-yl]-1,3-thiazol-2-yl)-2-[(tert-butoxycarbonyl)amino]propanoate(40 g, 61.01 mmol) in THF (300 mL) and H₂O (100 mL) were added LiOH(4.38 g, 183.05 mmol) dropwise at 0° C. The resulting mixture wasstirred for overnight at room temperature. The residue was acidified topH 6 with cone. HCl. The resulting mixture was extracted with EtOAc (500mL). The combined organic layers were washed with water (3×500 mL),dried over anhydrous Na₂SO₄. After filtration, the filtrate wasconcentrated under reduced pressure. This resulted in(2S)-2-[(tert-butoxycarbonyl)amino]-3-[4-[3-(3-hydroxy-2,2-dimethylpropyl)-2-iodo-1H-indol-5-yl]-1,3-thiazol-2-yl]propanoicacid (40 g, crude) as a yellow oil. The crude product was used in thenext step directly without further purification. LCMS (ESI): m/z [M+H]calc'd for C₂₄H₃₀IN₃O₅S 599.1.1; found 600.1.

Step 7

To a stirred solution of(2S)-2-[(tert-butoxycarbonyl)amino]-3-[4-[3-(3-hydroxy-2,2-dimethylpropyl)-2-iodo-1H-indol-5-yl]-1,3-thiazol-2-yl]propanoicacid (40 g, 66.72 mmol) and methyl (3S)-1,2-diazinane-3-carboxylate(28.86 g, 200.17 mmol) and HOBT (1.8 g, 13.35 mmol) and DIEA (172.47 g,1334.5 mmol) in DCM (350 mL) were added EDCl (31.98 g, 166.8 mmol)dropwise at 0° C. The resulting mixture was stirred for overnight atroom temperature under nitrogen atmosphere. The resulting mixture waswashed with water (1.5 L), dried over anhydrous Na₂SO₄. Afterfiltration, the filtrate was concentrated under reduced pressure. Theresidue was purified by silica gel column chromatography, eluted withPE/EtOAc (1:1) to afford methyl(3S)-1-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-[4-[3-(3-hydroxy-2,2-dimethylpropyl)-2-iodo-1H-indol-5-yl]-1,3-thiazol-2-yl]propanoyl]-1,2-diazinane-3-carboxylate(28 g, 43.9%) as a yellow oil. LCMS (ESI): m/z [M+H] calc'd forC₃₀H₄₀IN₅O₆S. 725.1.1; found 726.1.

Step 8

To a stirred solution of methyl(3S)-1-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-[4-[3-(3-hydroxy-2,2-dimethylpropyl)-2-iodo-1H-indol-5-yl]-1,3-thiazol-2-yl]propanoyl]-1,2-diazinane-3-carboxylate(28 g, 38.5 mmol) in THF (240 mL) were added LiOH (2.77 g, 115.7 mmol)in H₂O (80 mL) dropwise at 0° C. The resulting mixture was stirred for 2h at room temperature. The mixture was acidified to pH 6 with cone. HCl.The resulting mixture was extracted with EtOAc (300 mL). The combinedorganic layers were washed with water (3×300 mL), dried over anhydrousNa₂SO₄. After filtration, the filtrate was concentrated under reducedpressure. This resulted in(3S)-1-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-[4-[3-(3-hydroxy-2,2-dimethylpropyl)-2-iodo-1H-indol-5-yl]-1,3-thiazol-2-yl]propanoyl]-1,2-diazinane-3-carboxylicacid (25 g, crude) as a yellow oil. The crude product was used in thenext step directly without further purification. LCMS (ESI): m/z [M+H]calc'd for C₂₉H₃₈IN₅O₆S. 711.1; found 712.2.

Step 9

To a stirred solution of(3S)-1-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-[4-[3-(3-hydroxy-2,2-dimethylpropyl)-2-iodo-1H-indol-5-yl]-1,3-thiazol-2-yl]propanoyl]-1,2-diazinane-3-carboxylicacid (25 g, 35.13 mmol) and HOBT (23.74 g, 175.6 mmol) and DIPEA (136.21g, 1053.9 mmol) in DCM (2 L) were added EDCl (188.5 g, 983.6 mmol) inportions at 0° C. The resulting mixture was stirred for overnight atroom temperature under nitrogen atmosphere. The resulting mixture waswashed with water (6 L), dried over anhydrous Na₂SO₄. After filtration,the filtrate was concentrated under reduced pressure. The residue waspurified by silica gel column chromatography, eluted with PE/EtOAc (2:1)to afford tert-butyl((6³S,4S,Z)-1²-iodo-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(13 g, 45.88%) as a yellow solid. LCMS (ESI): m/z [M+H] calc'd forC₂₉H₃₆IN₅O₅S. 693.1; found 694.0.

Step 10

To a stirred mixture of tert-butyl((6³S,4S,Z)-1²-iodo-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(13 g, 18.7 mmol) and KOAc (6.44 g, 65.6 mmol) and s-Phos (2.31 g, 5.62mmol) in toluene (120 mL) were added Pd₂(dba)₃(2.06 g, 2.25 mmol) inportions at room temperature under argon atmosphere. To the stirredsolution were added 4,4,5,5-tetramethyl-1,3,2-dioxaborolane (17.99 g,140.5 mmol) dropwise at 0° C. under argon atmosphere. The resultingmixture was stirred for 3 h at 60° C. under argon atmosphere. Thereaction was quenched with sat. NH₄Cl (aq.) at 0° C. The resultingmixture was concentrated under vacuum. The resulting mixture wasextracted with EtOAc (200 mL). The combined organic layers were washedwith water (3×300 mL), dried over anhydrous Na₂SO₄. After filtration,the filtrate was concentrated under reduced pressure. The residue waspurified by silica gel column chromatography, eluted with PE/EtOAc (3:1)to afford tert-butyl((6³S,4S,Z)-10,10-dimethyl-5,7-dioxo-1²-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(10 g, 68.6% yield) as a yellow solid. LCMS (ESI): m/z [M+H] calc'd forC₃₅H₄₈BN₅O₇S. 693.3; found 694.4.

Intermediate 4. Synthesis of (S)-3-bromo-5-iodo-2-(1-methoxyethyl)pyridine

Step 1

To a stirred solution of 3-bromo-2-[(1S)-1-methoxyethyl]pyridine (80.00g, 370.24 mmol, 1.00 equiv) and bis(pinacolato)diboron (141.03 g, 555.3mmol, 1.50 equiv) in THF (320 mL) was added dtbpy (14.91 g, 55.5 mmol)and Chloro(1,5-cyclooctadiene)iridium(I) dimer (7.46 g, 11.1 mmol) underargon atmosphere. The resulting mixture was stirred for 16 h at 75° C.under argon atmosphere. The mixture was concentrated under reducedpressure. The resulting mixture was dissolved in EtOAc (200 mL) and themixture was adjusted to pH 10 with Na₂CO₃ (40 g) and NaOH (10 g) (mass4:1) in water (600 mL). The aqueous layer was extracted with EtOAc (800mL). The aqueous phase was acidified to pH=6 with HCl (6 N) toprecipitate the desired solid to afford5-bromo-6-[(1S)-1-methoxyethyl]pyridin-3-ylboronic acid (50 g, 52.0%yield) as a light-yellow solid. LCMS (ESI): m/z [M+H] calc'd forC₈H₁₁BBrNO₃ 259.0; found 260.0.

Step 2

To a stirred solution of5-bromo-6-[(1S)-1-methoxyethyl]pyridin-3-ylboronic acid (23.00 g, 88.5mmol) in ACN (230 mL) were added NIS (49.78 g, 221.2 mmol) at roomtemperature under argon atmosphere. The resulting mixture was stirredfor overnight at 80° C. under argon atmosphere. The resulting mixturewas concentrated under reduced pressure. The resulting mixture wasdissolved in DCM (2.1 L) and washed with Na₂S₂CO₃ (3×500 mL). Theorganic layer was dried over anhydrous Na2SO4. After filtration, thefiltrate was concentrated under reduced pressure. The residue waspurified by silica gel column chromatography to afford(S)-3-bromo-5-iodo-2-(1-methoxyethyl)pyridine (20 g, 66.0% yield). LCMS(ESI): m/z [M+H] calc'd for C₈H₉BrINO 340.9; found 341.7.

Intermediate 5. Synthesis of tert-butyl((6³S,4S,Z)-11-ethyl-1²-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate

Step 1

Into a 3 L 3-necked round-bottom flask purged and maintained with aninert atmosphere of argon, was placed3-bromo-5-iodo-2-[(1S)-1-methoxyethyl]pyridine (147 g, 429.8 mmol)benzyl piperazine-1-carboxylate (94.69 g, 429.8 mmol), Pd(OAc)₂ (4.83 g,21.4 mmol), BINAP (5.35 g, 8.6 mmol), Cs₂CO₃ (350.14 g, 1074.6 mmol),toluene (1 L). The resulting solution was stirred for overnight at 100°C. in an oil bath. The reaction mixture was cooled to 25° C. afterreaction completed. The resulting mixture was concentrated under reducedpressure. The residue was applied onto a silica gel column with ethylacetate/hexane (1:1). Removal of solvent under reduced pressure gavebenzyl(S)-4-(5-bromo-6-(1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylate(135 g, 65.1% yield) as a dark yellow solid. LCMS (ESI): m/z [M+H]calc'd for C₂₀H₂₄BrN₃O₃ 433.1; found 434.1.

Step 2

Into a 3-L 3-necked round-bottom flask purged and maintained with aninert atmosphere of argon, was placed benzyl4-[5-bromo-6-[(1S)-1-methoxyethyl]pyridin-3-yl]piperazine-1-carboxylate(135 g, 310.8 mmol), bis(pinacolato)diboron (86.82 g, 341.9 mmol),Pd(dppf)Cl₂ (22.74 g, 31.0 mmol), KOAc (76.26 g, 777.5 mmol), Toluene (1L). The resulting solution was stirred for 2 days at 90° C. in an oilbath. The reaction mixture was cooled to 25° C. The resulting mixturewas concentrated under vacuum. The residue was applied onto a neutralalumina column with ethyl acetate/hexane (1:3). Removal of solvent underreduced pressure gave benzyl(S)-4-(6-(1-methoxyethyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-3-yl)piperazine-1-carboxylate(167 g, crude) as a dark yellow solid. LCMS (ESI): m/z [M+H] calc'd forC₂₆H₃₆BN₃O₅ 481.3; found 482.1.

Step 3

Into a 3-L 3-necked round-bottom flask purged and maintained with aninert atmosphere of argon, was placed(S)-4-(6-(1-methoxyethyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-3-yl)piperazine-1-carboxylate(167 g, 346.9 mmol),5-bromo-3-[3-[(tert-butyldiphenylsilyl)oxy]-2,2-dimethylpropyl]-2-iodo-1H-indole(224.27 g, 346.9 mmol), Pd(dppf)Cl₂ (25.38 g, 34.6 mmol), dioxane (600mL), H₂O (200 mL), K₃PO₄ (184.09 g, 867.2 mmol), Toluene (200 mL). Theresulting solution was stirred for overnight at 70° C. in an oil bath.The reaction mixture was cooled to 25° C. after reaction completed. Theresulting mixture was concentrated under vacuum. The residue was appliedonto a silica gel column with ethyl acetate/hexane (1:1). Removal ofsolvent under reduced pressure gave benzyl(S)-4-(5-(5-bromo-3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-1H-indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylate(146 g, 48.1% yield) as a yellow solid. LCMS (ESI): m/z [M+H] calc'd forC₄₉H₅₇BrN₄O₄Si, 872.3; found 873.3.

Step 4

To a stirred mixture of benzyl(S)-4-(5-(5-bromo-3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-1H-indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylate(146 g, 167.0 mmol) and Cs₂CO₃ (163.28 g, 501.1 mmol) in DMF (1200 mL)was added C₂H₅I (52.11 g, 334.0 mmol) in portions at 0° C. under N₂atmosphere. The final reaction mixture was stirred at 25° C. for 12 h.Desired product could be detected by LCMS. The resulting mixture wasdiluted with EA (1 L) and washed with brine (3×1.5 L). The organiclayers were dried over anhydrous Na₂SO₄. After filtration, the filtratewas concentrated under reduced pressure to give benzyl(S)-4-(5-(5-bromo-3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-1-ethyl-1H-indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylate(143 g, crude) as a yellow solid that was used directly for next stepwithout further purification. LCMS (ESI): m/z [M+H] calc'd forC₅₁H₆₁BrN₄O₄Si, 900.4; found 901.4.

Step 5

To a stirred mixture of benzyl benzyl(S)-4-(5-(5-bromo-3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-1-ethyl-1H-indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylate(143 g, 158.5 mmol) in DMF (1250 mL) was added CsF (72.24 g, 475.5mmol). Then the reaction mixture was stirred at 60° C. for 2 days underN₂ atmosphere. Desired product could be detected by LCMS. The resultingmixture was diluted with EA (1 L) and washed with brine (3×1 L). Thenthe organic phase was concentrated under reduced pressure. The residuewas purified by silica gel column chromatography, eluted with PE/EA(1/3) to afford two atropisomers of benzyl(S)-4-(5-(5-bromo-1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-1H-indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylateA (38 g, 36% yield, RT=1.677 min in 3 min LCMS (0.1% FA)) and B (34 g,34% yield, RT=1.578 min in 3 min LCMS (0.1% FA)) both as yellow solid.LCMS (ESI): m/z [M+H] calc'd for C₃₅H₄₃BrN₄CO₄ 663.2; found 662.2.

Step 6

Into a 500-mL 3-necked round-bottom flask purged and maintained with aninert atmosphere of nitrogen, was placed benzyl(S)-4-(5-(5-bromo-1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-1H-indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylateA (14 g, 21.1 mmol), bis(pinacolato)diboron (5.89 g, 23.21 mmol),Pd(dppf)Cl₂ (1.54 g, 2.1 mmol), KOAc (5.18 g, 52.7 mmol), Toluene (150mL). The resulting solution was stirred for 5 h at 90° C. in an oilbath. The reaction mixture was cooled to 25° C. The resulting mixturewas concentrated under vacuum. The residue was purified by silica gelcolumn chromatography, eluted with PE/EA (1/3) to give benzyl(S)-4-(5-(1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylate(12 g, 76.0% yield) as a yellow solid. LCMS (ESI): m/z [M+H] calc'd forC₄₁H₅₅BN₄O₆ 710.4; found 711.3.

Step 7

Into a 250-mL round-bottom flask purged and maintained with an inertatmosphere of argon, was placed benzyl(S)-4-(5-(1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylate(10.8 g, 15.2 mmol), methyl(3S)-1-[(2S)-3-(4-bromo-1,3-thiazol-2-yl)-2-[(tert-butoxycarbonyl)amino]propanoyl]-1,2-diazinane-3-carboxylate(7.98 g, 16.7 mmol), Pd(dtbpf)Cl₂ (0.99 g, 1.52 mmol), K₃PO₄ (8.06 g,37.9 mmol), Toluene (60 mL), dioxane (20 mL), H₂O (20 mL). The resultingsolution was stirred for 3 h at 70° C. in an oil bath. The reactionmixture was cooled to 25° C. The resulting solution was extracted withEtOAc (2×50 mL) and concentrated under reduced pressure. The residue wasapplied onto a silica gel column with ethyl acetate/hexane (10:1).Removal of solvent to give methyl(S)-1-((S)-3-(4-(2-(5-(4-((benzyloxy)carbonyl)piperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-1H-indol-5-yl)thiazol-2-yl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylate(8 g, 50.9% yield) as a yellow solid. LCMS (ESI): m/z [M+H] calc'd forC₅₂H₆₈N₈O₉S. 980.5; found 980.9.

Step 8

To a stirred mixture of methyl(S)-1-((S)-3-(4-(2-(5-(4-((benzyloxy)carbonyl)piperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-1H-indol-5-yl)thiazol-2-yl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylate(12 g, 12.23 mmol) in THF (100 mL)/H₂O (100 mL) was added LiOH (2.45 g,61.1 mmol) under N₂ atmosphere and the resulting mixture was stirred for2 h at 25° C. Desired product could be detected by LCMS. THF wasconcentrated under reduced pressure. The pH of aqueous phase wasacidified to 5 with HCL (1N) at 0° C. The aqueous layer was extractedwith DCM (3×100 ml). The organic phase was concentrated under reducedpressure to give(S)-1-((S)-3-(4-(2-(5-(4-((benzyloxy)carbonyl)piperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-1H-indol-5-yl)thiazol-2-yl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylicacid (10 g, 84.5% yield) as a light yellow solid. LCMS (ESI): m/z [M+H]calc'd for C₅₁H₆₆N₈O₉S. 966.5; found 967.0.

Step 9

Into a 3-L round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed(S)-1-((S)-3-(4-(2-(5-(4-((benzyloxy)carbonyl)piperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-1H-indol-5-yl)thiazol-2-yl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylicacid (18 g, 18.61 mmol), ACN (1.8 L), DIEA (96.21 g, 744.4 mmol), EDCl(107.03 g, 558.3 mmol), HOBT (25.15 g, 186.1 mmol). The resultingsolution was stirred for overnight at 25° C. The resulting mixture wasconcentrated under vacuum after reaction completed. The resultingsolution was diluted with DCM (1 L). The resulting mixture was washedwith HCl (3×1 L, 1N aqueous). The resulting mixture was washed withwater (3×1 L). Then the organic layer was concentrated, the residue wasapplied onto a silica gel column with ethyl acetate/hexane (1:1).Removal of solvent under reduced pressure gave benzyl4-(5-((6³S,4S,Z)-4-((tert-butoxycarbonyl)amino)-1¹-ethyl-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-1²-yl)-6-((S)-1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylate(10.4 g, 54.8% yields) as a light yellow solid. LCMS (ESI): m/z [M+H]calc'd for C₅₁H₆₄N₈O₈S. 948.5; found 949.3.

Step 10

Into a 250-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed benzyl4-(5-((6³S,4S,Z)-4-((tert-butoxycarbonyl)amino)-1¹-ethyl-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-1²-yl)-6-((S)-1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylate(10.40 g, 10.9 mmol), Pd(OH)₂/C (5 g, 46.9 mmol), MeOH (100 mL). Theresulting solution was stirred for 3 h at 25° C. under 2 atm H₂atmosphere. The solids were filtered out and the filter cake was washedwith MeOH (3×100 mL). Then combined organic phase was concentrated underreduced pressure to give tert-butyl((6³S,4S,Z)-11-ethyl-1²-(2-((S)-1-methoxyethyl)-5-(piperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(8.5 g, 90.4% yield) as a light yellow solid. LCMS (ESI): m/z [M+H]calc'd for C₄₃H₅₈N₈O₆S. 814.4; found 815.3.

Step 11

Into a 1000-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed tert-butyl ((6³S,4S,Z)-1f-ethyl-1²-(2-((S)-1-methoxyethyl)-5-(piperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(8.5 g, 10.4 mmol), MeOH (100 mL), AcOH (1.88 g, 31.2 mmol) and stirredfor 15 mins. Then HCHO (1.88 g, 23.15 mmol, 37% aqueous solution) andNaBH₃CN (788 mg, 12.5 mmol) was added at 25° C. The resulting solutionwas stirred for 3 h at 25° C. The resulting mixture was quenched with100 mL water and concentrated under reduced pressure to remove MeOH. Theresulting solution was diluted with 300 mL of DCM. The resulting mixturewas washed with water (3×100 mL). Removal of solvent gave tert-butyl((6³S,4S,Z)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(8.2 g, 90.1% yield) as a yellow solid. LCMS (ESI): m/z [M+H] calc'd forC₄₄H₆₀N₈O₆S. 828.4; found 829.3.

Example A120.(1S,2S)—N-((6³S,4S,Z)-1¹-ethyl-1²-(5-((S)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2-methylcyclopropane-1-carboxamide

Step 1

To a stirred solution of 3-bromo-5-iodo-2-[(1S)-1-methoxyethyl]pyridine(1 g, 2.92 mmol) (S)-octahydropyrazino[2,1-c][1,4]oxazine (498.9 mg, 3.1mmol) and Potassium tert-butoxide (656.25 mg, 5.8 mmol) in Toluene (15mL) were added Pd₂(dba)₃ (53.55 mg, 0.06 mmol) and XantPhos (169.2 mg,0.29 mmol) at room temperature under nitrogen atmosphere. The resultingmixture was stirred for 3 h at 100° C. under nitrogen atmosphere. Aftercompletion of reaction, the solution was concentrated under reducedpressure. The residue was purified by reverse flash chromatography(conditions: column, C₁₈ silica gel; mobile phase, ACN in water (0.05%TFA), 0% to 100% gradient in 40 min; detector, UV 254 nm) to afford(S)-8-(5-bromo-6-((S)-1-methoxyethyl)pyridin-3-yl)octahydropyrazino[2,1-c][1,4]oxazine(670 mg, 57.2% yield) as a yellow solid. LCMS (ESI): m/z [M+H] calc'dfor C₁₅H₂₂BrN₃O₂ 355.1; found 356.1.

Step 2

To a mixture of(S)-8-(5-bromo-6-((S)-1-methoxyethyl)pyridin-3-yl)octahydropyrazino[2,1-c][1,4]oxazine(670 mg, 1.88 mmol), Intermediate 3 (1.56 g, 2.26 mmol) and K₂CO₃(779.74 mg, 5.6 mmol) in Toluene (9 mL), H₂O (3 mL) and 1,4-dioxane (3mL) was added Pd(dppf)Cl₂ (137.61 mg, 0.19 mmol) at room temperatureunder nitrogen atmosphere. The resulting mixture was stirred forovernight at 65° C. under nitrogen atmosphere. After completion ofreaction, the solution was concentrated under reduced pressure. Theresidue was purified by reverse flash chromatography (conditions:column, C₁₈ silica gel; mobile phase, ACN in water (0.05% TFA), 0% to100% gradient in 30 min; detector, UV 254 nm) to afford tert-butyl((6³S,4S,Z)-1²-(5-((S)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(1.4 g, 88.3% yield) as a white solid. LCMS (ESI): m/z [M+H] calc'd forC₄₄H₅₈N₈O₇S. 842.4; found 843.2.

Step 3

To a stirred mixture of tert-butyl ((6³S,4S,Z)-1²-(5-((S)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(1.4 g, 1.66 mmol) and Cs₂CO₃ (1.62 g, 4.97 mmol) in DMF (10 mL) wasadded ethyl iodide (0.39 g, 2.5 mmol) dropwise at 0° C. The resultingmixture was stirred for 2 h at room temperature. After completion ofreaction, the solution was concentrated under reduced pressure. Theresidue was purified by reverse flash chromatography (conditions:column, C18 silica gel; mobile phase, ACN in water (0.05% TFA), 0% to100% gradient in 30 min; detector, UV 254 nm) to afford tert-butyl((6³S,4S,Z)-1¹-ethyl-1²-(5-((S)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(800 mg, 49.7% yield) as a brown yellow solid. LCMS (ESI): m/z [M+H]calc'd for C₄₆H₆₂N₈O₇S. 870.4; found 871.2.

Step 4

Into a 50 mL round-bottom flask were added tert-butyl ((6³ S, 4S,Z)-1¹-ethyl-1²-(5-((S)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(250 mg, 0.29 mmol) and HCl (4M in 1,4-dioxane, 10 mL) at 0° C. Theresulting mixture was stirred for 1 h at room temperature. The resultingmixture was concentrated. The resulting mixture was diluted with 30 mLof dichloromethane and 20 mL saturated NaHCO₃ aqueous solution. Theorganic phase was washed twice with 30 mL brine. Removal of solventunder reduced pressure resulted in(6³S,4S,Z)-4-amino-1′-ethyl-1²-(5-((S)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-5,7-dione(170.00 mg, crude) as a brown solid. LCMS (ESI): m/z [M+H] calc'd forC₄₁H₅₄N₈O₅S. 770.4; found 771.2.

Step 5

To a stirred solution of(6³S,4S,Z)-4-amino-1′-ethyl-1²-(5-((S)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-5,7-dione(170 mg, 0.22 mmol) in DMF (8 mL) were added DIEA (2.8 g, 22 mmol),(1S,2S)-2-methylcyclopropane-1-carboxylic acid (33 mg, 0.33 mmol) andHATU (125 mg, 0.33 mmol) at 0° C. The resulting mixture was stirred for2 h at room temperature. After completion of reaction, the solution wasconcentrated under reduced pressure. The crude product was purified byPrep-HPLC to afford 50 mg racemated product. The racemate was purifiedby Prep-CHIRAL-HPLC with the following conditions (Column: CHIRAL ARTCellulose-SB, 2*25 cm, 5 um; Mobile Phase A: MtBE (10 mM NH₃-MeOH),Mobile Phase B: EtOH; Flow rate: 20 mL/min; Gradient: 50% B to 50% B in7 min; 275/210 nm) to afford two atropisomers of(1S,2S)—N-((6³S,4S,Z)-1¹-ethyl-1²-(5-((S)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2-methylcyclopropane-1-carboxamide(as single atropisomer) (10.7 mg, 5.1% yield) and (6 mg, 3.03%) both aswhite solid. LCMS (ESI): m/z [M+H] calc'd for C₄₆H₆₀N₈O₆S 852.3; found853.5. Isomer 1. ¹H NMR (400 MHz, DMSO-d6) δ 8.53 (d, 2H), 8.46 (d, 1H),7.79 (s, 1H), 7.71 (d, 1H), 7.51 (d, 1H), 7.38 (d, 1H), 5.55 (t, 1H),5.05 (d, 1H), 4.22 (t, 2H), 3.99-3.80 (m, 4H), 3.82-3.59 (m, 5H), 3.54(d, 2H), 3.39 (d, 1H), 3.14 (t, 2H), 3.07 (s, 3H), 2.99 (s, 1H), 2.81(t, 3H), 2.67 (d, 1H), 2.44-2.34 (m, 2H), 2.30 (s, 1H), 2.21-2.07 (m,2H), 1.80 (s, 2H), 1.51 (s, 2H), 1.21 (d, 4H), 1.15-0.93 (m, 7H), 0.87(s, 3H), 0.65 (m, 2H), 0.52 (s, 4H). Isomer 2. ¹H NMR (400 MHz, DMSO-d6)δ 8.54-8.39 (m, 3H), 7.79 (s, 1H), 7.75-7.67 (m, 1H), 7.55 (d, 1H), 7.22(d, 1H), 5.56 (t, 1H), 5.07 (d, 1H), 4.34-4.09 (m, 5H), 3.83-3.62 (m,4H), 3.55 (d, 3H), 3.21 (s, 3H), 3.14 (d, 2H), 2.94-2.64 (m, 5H),2.46-2.36 (m, 2H), 2.32-2.15 (m, 3H), 2.08 (d, 1H), 1.79 (s, 2H), 1.49(s, 2H), 1.33 (d, 3H), 1.25 (d, 1H), 1.06 (s, 4H), 0.90 (d, 7H), 0.54(d, 1H), 0.34 (s, 3H).

Example A14.N-[(7S,13S,19M)-21-ethyl-20-{2-[(1S)-1-methoxyethyl]pyridin-3-yl}-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1²,⁵.1⁹,¹³.0²²,²⁶]octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]azetidine-3-carboxamide

Step 1

To a stirred solution of(6³S,4S,Z)-4-amino-1′-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-5,7-dione(450.00 mg, 0.71 mmol, 1.00 equiv) and1-(tert-butoxycarbonyl)azetidine-3-carboxylic acid (215.3 mg, 1.07 mmol)in DMF (5.00 mL) were added DIEA (460.99 mg, 3.5 mmol) and HATU (379.7mg, 1 mmol) in portions at room temperature under N₂ atmosphere untilthe reaction was complete by LCMS. The resulting mixture was extractedwith EtOAc (3×20 mL) and the combined organic layers were washed withbrine (2×20 mL), dried over anhydrous Na₂SO₄. After filtration, thefiltrate was concentrated under reduced pressure and the resultingresidue was purified by silica gel column chromatography to affordtert-butyl3-(((6³S,4S,Z)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamoyl)azetidine-1-carboxylate(520 mg, 90%) as a yellow solid. LCMS (ESI): m/z [M+H] calc'd forC₄₃H₅₅N₇O₇S ESI-MS 813.4; found: 814.4.

Step 2

To a solution of tert-butyl3-(((6³S,4S,Z)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamoyl)azetidine-1-carboxylate(170.00 mg, 0.21 mmol) in DCM (1.6 mL) was added TFA (0.4 mL, 5.3 mmol)dropwised at 0° C. It was stirred for 2 h at room temperature under N₂atmosphere and then concentrated under reduced pressure. The crudeproduct was purified by Prep-HPLC to affordN-[(7S,13S,19M)-21-ethyl-20-{2-[(1S)-1-methoxyethyl]pyridin-3-yl}-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetraazapentacyclo[17.5.2.1²,⁵.1⁹,¹³.0²²,²⁶]octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]azetidine-3-carboxamide (44.7 mg, 30% yield) asa white solid. LCMS (ESI): m/z [M+H] calc'd for C₃₈H₄₇N₇O₅S. 713.3;found 714.1.

Example A99.(1S,2S)—N-((6³S,4S,Z)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2-phenylcyclopropane-1-carboxamide

To a solution of tert-butyl((6³S,4S,Z)-11-ethyl-1²-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(50.0 mg, 0.069 mmol), (1S,2S)-2-phenylcyclopropane-1-carboxylic acid(16.69 mg, 0.103 mmol), and DIPEA (44.32 mg, 0.343 mmol) in DMF (0.50mL) at 0° C. was added HATU (78.24 mg, 0.206 mmol). The resultingmixture was warmed to room temperature and stirred for 3 h. The crudeproduct was purified by prep-HPLC to give(1S,2S)—N-((6³S,4S,Z)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2-phenylcyclopropane-1-carboxamide(34 mg, 51% yield) as an off-white solid. LCMS (ESI): m/z [M+H] calc'dfor C₄₉H₆₀N₈O₅S. 873.5; found 874.1.

Example A121. Synthesis of(1S,2S)—N-((6³S,4S,Z)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-(4-(2-methoxyethyl)piperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2-methylcyclopropane-1-carboxamide

Step 1

A mixture of benzyl4-(5-((6³S,4S,Z)-4-((tert-butoxycarbonyl)amino)-1¹-ethyl-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-12-yl)-6-((S)-1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylate(380 mg, 0.4 mmol) in EtOAc (10 mL) was added Pd(OH)₂/C (600 mg, 20 mol%) was hydrogenated at rt overnight. The mixture was filtered through apad of Celite® and the filtrate was concentrated under reduced pressureto give tert-butyl((6³S,4S,Z)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-(piperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate (310 mg, crude) as a solid.LCMS (ESI): m/z [M+H] calc'd for C₄₃H₅₈N₈O₆S. 814.4; found 815.5.

Step 2

To a mixture of tert-butyl((6³S,4S,Z)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-(piperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate (300 mg, 0.37 mmol) and1-bromo-2-methoxyethane (56 mg, 0.41 mmol) in MeCN (10 mL) at rt wasadded KI (61 mg, 0.37 mmol) and K₂CO₃ (51 mg, 0.37 mmol) in portions.The mixture was heated to 60° C. and stirred for 2 h, then diluted withH₂O (5 mL). The residue was purified by preparative-HPLC to givetert-butyl((6³S,4S,Z)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-(4-(2-methoxyethyl)piperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(310 mg, 97% yield) as a solid. LCMS (ESI): m/z [M+H] calc'd forC₄₆H₆₄N₈O₇S. 872.5; found 873.6.

Step 3

A mixture of tert-butyl((6³S,4S,Z)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-(4-(2-methoxyethyl)piperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(300 mg, 0.34 mmol) in 4 M HCl in 1,4-dioxane, (10 mL) was stirred at rtfor 1 h, then concentrated under reduced pressure to give(6³S,4S,Z)-4-amino-1′-ethyl-1²-(2-((S)-1-methoxyethyl)-5-(4-(2-methoxyethyl)piperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-5,7-dioneHCl salt (315 mg, crude) as a solid, which was used directly in the nextstep without further purification. LCMS (ESI): m/z [M+H] calc'd forC₄₁H₅₆N₈O₅S. 772.4; found 773.3.

Step 4

A mixture of(6³S,4S,Z)-4-amino-1′-ethyl-1²-(2-((S)-1-methoxyethyl)-5-(4-(2-methoxyethyl)piperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-5,7-dioneHCl salt (300 mg, 0.39 mmol) and(1S,2S)-2-methylcyclopropane-1-carboxylic acid (97 mg, 0.97 mmol) in DMF(5 mL) at 0° C. was added DIPEA (1.00 g, 7.77 mmol) dropwise, then COMU(249 mg, 0.58 mmol) in portions. The mixture was allowed to warm to rtand stirred for 2 h, then concentrated under reduced pressure and theresidue was purified by preparative-HPLC to give(1S,2S)—N-((6³S,4S,Z)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-(4-(2-methoxyethyl)piperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2-methylcyclopropane-1-carboxamide(178 mg, 54% yield) as a solid. LCMS (ESI): m/z [M+H] calc'd forC₄₆H₆₂N₈O₆S. 854.5; found 855.5; ¹H NMR (400 MHz, DMSO-d₆) δ 8.55-8.46(m, 2H), 7.80 (d, J=3.1 Hz, 1H), 7.77-7.70 (m, 1H), 7.57 (d, J=8.6 Hz,1H), 7.40 (d, J=2.8 Hz, 1H), 5.55 (d, J=9.2 Hz, 1H), 4.37-4.09 (m, 6H),4.00 (s, 2H), 3.83-3.71 (m, 2H), 3.57 (s, 3H), 3.38 (t, J=4.8 Hz, 2H),3.32 (d, J=2.5 Hz, 4H), 3.22 (s, 7H), 3.20-3.11 (m, 1H), 2.94 (d, J=14.4Hz, 1H), 2.76 (t, J=11.4 Hz, 1H), 2.44 (d, J=14.2 Hz, 1H), 2.07 (d,J=12.0 Hz, 1H), 1.80 (s, 2H), 1.60-1.47 (m, 2H), 1.34 (d, J=6.1 Hz, 3H),1.07 (d, J=1.8 Hz, 4H), 0.95-0.82 (m, 7H), 0.55 (d, J=7.4 Hz, 1H), 0.35(s, 3H).

Example A157. Synthesis of(1S,2S)—N-((6³S,4S,Z)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-((R)-octahydro-2H-pyrido[1,2-a]pyrazin-2-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2-methylcyclopropane-1-carboxamide

Step 1

5-[(9aR)-octahydropyrido[1,2-a]pyrazin-2-yl]-3-bromo-2-[(1S)-1-methoxyethyl]pyridinewas synthesized in a manner similar to(S)-8-(5-bromo-6-((S)-1-methoxyethyl)pyridin-3-yl)octahydropyrazino[2,1-c][1,4]oxazineexcept (S)-octahydropyrazino[2,1-c][1,4]oxazine was substituted with(S)-octahydropyrazino[2,1-c][1,4]oxazine (1.5 g, 60% yield) as a solid.LCMS (ESI): m/z [M+H] calc'd for C₁₆H₂₄BrN₃O 353.1; found 354.1.

Step 2

Tert-butyl((6³S,4S,Z)-1²-(2-((S)-1-methoxyethyl)-5-((R)-octahydro-2H-pyrido[1,2-a]pyrazin-2-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamatewas synthesized in a manner similar to tert-butyl((6³S,4S,Z)-1²-(5-((S)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate except(S)-8-(5-bromo-6-((S)-1-methoxyethyl)pyridin-3-yl)octahydropyrazino[2,1-c][1,4]oxazinewas substituted with5-[(9aR)-octahydropyrido[1,2-a]pyrazin-2-yl]-3-bromo-2-[(1S)-1-methoxyethyl]pyridineand K₂CO₃ was substituted with K₃PO₄ to give (800 mg, 83% yield) as anoil. LCMS (ESI): m/z [M+H] calc'd for C₄₅H₆₀N₈O₆S. 840.4; found 841.4.

Step 3

Tert-butyl((6³S,4S,Z)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-((R)-octahydro-2H-pyrido[1,2-a]pyrazin-2-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamatewas synthesized in a manner similar to tert-butyl((6³S,4S,Z)-1¹-ethyl-1²-(5-((S)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,66-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamateexcept tert-butyl((6³S,4S,Z)-1²-(5-((S)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamatewas substituted with tert-butyl((6³S,4S,Z)-1²-(2-((S)-1-methoxyethyl)-5-((R)-octahydro-2H-pyrido[1,2-a]pyrazin-2-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamateto give (220 mg, 27% yield) as a solid. LCMS (ESI): m/z [M+H] calc'd forC₄₇H₆₄N₈O₆S 868.5; found 869.5.

Step 4

A mixture of tert-butyl((6³S,4S,Z)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-((R)-octahydro-2H-pyrido[1,2-a]pyrazin-2-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(220 mg, 0.25 mmol) in 1,4-dioxane (2 mL) at 0° C. was added 4M HCl in1,4-dioxane (1 mL). The mixture was stirred at 0° C. for 1 h thenconcentrated under reduced pressure to give(6³S,4S,Z)-4-amino-1′-ethyl-1²-(2-((S)-1-methoxyethyl)-5-((R)-octahydro-2H-pyrido[1,2-a]pyrazin-2-yl)pyridin-3-yl)-10,10-dimethyl-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-5,7-dione(220 mg, crude) as a solid. LCMS (ESI): m/z [M+H] calc'd forC₄₂H₅₆N₈O₄S. 768.4; found 769.4.

Step 5

(1S,2S)—N-((6³S,4S,Z)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-((R)-octahydro-2H-pyrido[1,2-a]pyrazin-2-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2-methylcyclopropane-1-carboxamidewas synthesized in a manner similar to(1S,2S)—N-((6³S,4S,Z)-1¹-ethyl-1²-(5-((S)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2-methylcyclopropane-1-carboxamideexcept(6³S,4S,Z)-4-amino-1¹-ethyl-1²-(5-((S)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-5,7-dioneHCl salt was substituted with(6³S,4S,Z)-4-amino-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-((R)-octahydro-2H-pyrido[1,2-a]pyrazin-2-yl)pyridin-3-yl)-10,10-dimethyl-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-5,7-dioneto give (13 mg, 8% yield) as a solid. LCMS (ESI): m/z [M+H] calc'd forC₄₇H₆₂N₈O₅S. 850.5; found 851.6; ¹H NMR (400 MHz, DMSO-d₆) δ 8.53-8.41(m, 3H), 7.79 (s, 1H), 7.72-7.70 (m, 1H), 7.55-7.30 (m, 1H), 7.20-7.10(m, 1H), 5.56-5.46 (m, 1H), 5.08-5.00 (m, 1H), 4.39-4.04 (m, 5H),3.72-7.62 (m, 2H), 3.57-3.47 (m, 2H), 3.21-3.11 (m, 3H), 3.15-3.08 (m,1H), 2.94 (m, 1H), 2.79-2.69 (m, 4H), 2.45-2.35 (m, 3H), 2.24-2.22 (m,1H), 2.08-2.00 (m, 1H), 2.01-1.88 (m, 2H), 1.81-1.65 (m, 3H), 1.59 (d,J=12.1 Hz, 2H), 1.54-1.38 (m, 2H), 1.33-1,30m, 3H), 1.28-1.12 (m, 3H),1.06-0.86 (m, 4H), 0.96-0.79 (m, 6H), 0.55-0.50 (m, 1H), 0.34 (s, 3H).

Example A214. Synthesis of(1S,2S)—N-((6³S,4S,Z)-1¹-(2-cyanopropan-2-yl)-10,10-dimethyl-1²-(2-(4-methylpiperazin-1-yl)pyridin-4-yl)-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2-methylcyclopropane-1-carboxamide

Step 1

A mixture of benzyl 4-(4-bromopyridin-2-yl)piperazine-1-carboxylate(8.09 g, 21.5 mmol), bis(pinacolato)diboron (8.19 g, 32.3 mmol), KOAc(6.33 g, 64.5 mmol), Pd(dppf)Cl₂ (0.79 g, 1.1 mmol) in toluene (100 mL)under an atmosphere of Ar was heated to 90° C. and stirred for 2 h. Themixture was concentrated under vacuum, H2O (50 mL) was added to theresidue and the mixture was extracted with EtOAc (2×100 mL). Thecombined organic layers were washed with brine (2×100 mL), dried,filtered and the filtrate concentrated under reduced pressure to givebenzyl4-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-yl]piperazine-1-carboxylate(9.2 g, 100% yield) as a solid. LCMS (ESI): m/z [M+H] calc'd forC₂₃H₃₀BN₃O₄ 423.2; found 424.2.

Step 2

A mixture of benzyl4-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-yl]piperazine-1-carboxylate(5.00 g, 11.8 mmol),5-bromo-3-[3-[(tert-butyldiphenylsilyl)oxy]-2,2-dimethylpropyl]-2-iodo-1H-indole(7.64 g, 11.8 mmol), Pd(dppf)Cl₂ (0.86 g, 1.2 mmol), K₂CO₃ (6.27 g, 45.4mmol) in toluene (45 mL), 1,4-dioxane (15 mL), H₂O (15 mL) under anatmosphere of N₂ was heated to 70° C. and stirred for 2 h. H₂O (50 mL)was added and the mixture was extracted with EtOAc (2×50 mL). Thecombined organic layers were dried over anhydrous Na₂SO₄ and filtered.The filtrate was concentrated under reduced pressure and the residue waspurified by silica gel column chromatography to give benzyl4-[4-(5-bromo-3-[3-[(tert-butyldiphenylsilyl)oxy]-2,2-dimethylpropyl]-1H-indol-2-yl)pyridin-2-yl]piperazine-1-carboxylate(4.9 g, 51% yield) as solid. LCMS (ESI): m/z [M+H] calc'd forC₄₆H₅₁BrN₄O₃Si, 814.3; found 815.4.

Step 3

A mixture of benzyl4-[4-(5-bromo-3-[3-[(tert-butyldiphenylsilyl)oxy]-2,2-dimethylpropyl]-1H-indol-2-yl)pyridin-2-yl]piperazine-1-carboxylate(4.5 g, 5.5 mmol), 2-bromo-2-methylpropanamide (2.75 g, 16.6 mmol),K₃PO₄ (2.34 g, 11.0 mmol), NaOH (0.57 g, 14.3 mmol), PhbP (0.29 g, 1.1mmol), copper bromide-dimethyl sulfide (0.23 g, 1.1 mmol) in toluene (50mL) under an atmosphere of N₂ was heated to 45° C. and stirred for 2days. H₂O (50 mL) was added and the mixture was extracted with EtOAc(2×50 mL). The combined organic layers were washed with brine (2×50 mL),dried over Na₂SO₄ and filtered. The filtrate was concentrated underreduced pressure and the residue was purified by silica gel columnchromatography to give benzyl4-[4-(5-bromo-3-[3-[(tert-butyldiphenylsilyl)oxy]-2,2-dimethylpropyl]-1-(1-carbamoyl-1-methylethyl)indol-2-yl)pyridin-2-yl]piperazine-1-carboxylate(1.5 g, 30% yield) as a solid. LCMS (ESI): m/z [M+H] calc'd forC₅₀H₅₈BrN₅O₄Si, 899.3; found 900.4.

Step 4

A mixture of benzyl4-[4-(5-bromo-3-[3-[(tert-butyldiphenylsilyl)oxy]-2,2-dimethylpropyl]-1-(1-carbamoyl-1-methylethyl)indol-2-yl)pyridin-2-yl]piperazine-1-carboxylate(1.40 g, 1.6 mmol), Et₃N (0.47 g, 4.7 mmol) and TFAA (0.65 g, 3.1 mmol)in DCM (20 mL), 3 equiv) was stirred at rt for 2 h. H₂O (20 mL) wasadded and the mixture was extracted with DCM (2×20 mL). The combinedorganic layers were washed with brine (2×50 mL), dried over Na₂SO₄ andfiltered. The filtrate was concentrated under reduced pressure to givebenzyl4-[4-(5-bromo-3-[3-[(tert-butyldiphenylsilyl)oxy]-2,2-dimethylpropyl]-1-(1-cyano-1-methylethyl)indol-2-yl)pyridin-2-yl]piperazine-1-carboxylate(1.3 g, 95% yield) as a solid. LCMS (ESI): m/z [M+H] calc'd forC₅₀H₅₆BrN₅O₃Si, 881.3; found 882.4.

Step 5

A mixture of benzyl4-[4-(5-bromo-3-[3-[(tert-butyldiphenylsilyl)oxy]-2,2-dimethylpropyl]-1-(1-cyano-1-methylethyl)indol-2-yl)pyridin-2-yl]piperazine-1-carboxylate(1.50 g, 1.7 mmol), bis(pinacolato)diboron (5.21 g, 20.5 mmol),Pd₂(dba)₃ (0.38 g, 0.4 mmol), KOAc (1.21 g, 12.3 mmol), X-Phos (0.20 g,0.4 mmol) in 1,4-dioxane (25 mL) under an atmosphere of N₂ was heated to110° C. and stirred for 2 h. H₂O (25 mL) was added and the mixture wasextracted with EtOAc (2×25 mL). The combined organic layers were washedwith brine (2×25 mL), dried over Na₂SO₄ and filtered. The filtrate wasconcentrated under reduced pressure and the residue was purified bysilica gel column chromatography to give benzyl4-[4-(3-[3-[(tert-butyldiphenylsilyl)oxy]-2,2-dimethylpropyl]-1-(1-cyano-1-methylethyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indol-2-yl)pyridin-2-yl]piperazine-1-carboxylate(1.6 g, 94% yield) as an oil. LCMS (ESI): m/z [M+H] calc'd forC₅₆H₆₈BN₅O₅Si, 929.5; found 930.4.

Step 6

A mixture of benzyl4-[4-(3-[3-[(tert-butyldiphenylsilyl)oxy]-2,2-dimethylpropyl]-1-(1-cyano-1-methylethyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indol-2-yl)pyridin-2-yl]piperazine-1-carboxylate(1.60 g, 1.7 mmol), methyl(3S)-1-[(2S)-3-(4-bromo-1,3-thiazol-2-yl)-2-[(tert-butoxycarbonyl)amino]propanoyl]-1,2-diazinane-3-carboxylate(0.82 g, 1.7 mmol), K₂C₀₃ (0.79 g, 5.8 mmol), Pd(dppf)Cl₂ (0.13 g, 0.17mmol) in toluene (12 mL), 1,4-dioxane (4 mL) and H₂O (4 mL) under anatmosphere of N₂ was heated to 70° C. and stirred for 5 h. H₂O (30 mL)was added and the mixture was extracted with EtOAc (2×30 mL). Thecombined organic layers were washed with brine (2×30 mL), dried overNa₂SO₄ and filtered. The filtrate was concentrated under reducedpressure and the residue was purified by silica gel columnchromatography to give benzyl4-[4-(5-[2-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-[(3S)-3-(methoxycarbonyl)-1,2-diazinan-1-yl]-3-oxopropyl]-1,3-thiazol-4-yl]-3-[3-[(tert-butyldiphenylsilyl)oxy]-2,2-dimethylpropyl]-1-(1-cyano-1-methylethyl)indol-2-yl)pyridin-2-yl]piperazine-1-carboxylate(650 mg, 31% yield) as an oil. LCMS (ESI): m/z [M+H] calc'd forC₆₇H₈₁N₉O₈SSi 1199.6; found 1200.5.

Step 7

To a mixture of benzyl4-[4-(5-[2-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-[(3S)-3-(methoxycarbonyl)-1,2-diazinan-1-yl]-3-oxopropyl]-1,3-thiazol-4-yl]-3-[3-[(tert-butyldiphenylsilyl)oxy]-2,2-dimethylpropyl]-1-(1-cyano-1-methylethyl)indol-2-yl)pyridin-2-yl]piperazine-1-carboxylate(650 mg, 0.54 mmol) in THF (15 mL) under an atmosphere of N2 was addedTBAF (1.42 g, 5.4 mmol). The mixture was stirred at rt overnight thenthe mixture adjusted to pH ˜6 with 1M HCl and extracted with EtOAc (2×50mL). The combined organic layers were washed with brine (2×50 mL), driedover Na₂SO₄ and filtered. The filtrate was concentrated under reducedpressure and the residue was purified by silica gel columnchromatography to give(3S)-1-[(2S)-3-[4-[2-(2-[4-[(benzyloxy)carbonyl]piperazin-1-yl]pyridin-4-yl)-1-(1-cyano-1-methylethyl)-3-(3-hydroxy-2,2-dimethylpropyl)indol-5-yl]-1,3-thiazol-2-yl]-2-[(tert-butoxycarbonyl)amino]propanoyl]-1,2-diazinane-3-carboxylicacid (370 mg, 72% yield) as a solid. LCMS (ESI): m/z [M+H] calc'd forC₅₀H₆₁N₉O₈S. 947.4; found 948.5.

Step 8

A mixture of(3S)-1-[(2S)-3-[4-[2-(2-[4-[(benzyloxy)carbonyl]piperazin-1-yl]pyridin-4-yl)-1-(1-cyano-1-methylethyl)-3-(3-hydroxy-2,2-dimethylpropyl)indol-5-yl]-1,3-thiazol-2-yl]-2-[(tert-butoxycarbonyl)amino]propanoyl]-1,2-diazinane-3-carboxylicacid (370 mg, 0.39 mmol), DIPEA (1.51 g, 11.7 mmol), HOBT (264 mg, 1.95mmol), EDCl (2.09 g, 10.9 mmol) in DCM (370 mL) was stirred at rtovernight. H₂O (100 mL) was added and the mixture was extracted withEtOAc (2×100 mL). The combined organic layers were washed with brine(2×100 mL), dried over Na₂SO₄, filtered and the filtrate concentratedunder reduced pressure to give benzyl4-(4-((6³S,4S,Z)-4-((tert-butoxycarbonyl)amino)-1¹-(2-cyanopropan-2-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-12-yl)pyridin-2-yl)piperazine-1-carboxylate(187 mg, 52% yield) as a solid. LCMS (ESI): m/z [M+H] calc'd forC₅₀H₅₉N₉O₇S. 929.4; found 930.8.

Step 9

A mixture of benzyl4-(4-((6³S,4S,Z)-4-((tert-butoxycarbonyl)amino)-1¹-(2-cyanopropan-2-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-12-yl)pyridin-2-yl)piperazine-1-carboxylate(170 mg, 0.18 mmol), paraformaldehyde (165 mg, 1.8 mmol), Pd(OH)₂/C (170mg, 1.2 mmol) in MeOH (25 mL) was stirred under an atmosphere of H₂overnight. The mixture was filtered and the filtrate was concentratedunder reduced pressure to give tert-butyl((6³S,4S,Z)-1¹-(2-cyanopropan-2-yl)-10,10-dimethyl-1²-(2-(4-methylpiperazin-1-yl)pyridin-4-yl)-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(110 mg, 74% yield) as a solid. LCMS (ESI): m/z [M+H] calc'd forC₄₃H₅₅N₉O₅S. 809.4; found 810.9.

Step 10

A mixture of tert-butyl((6³S,4S,Z)-1¹-(2-cyanopropan-2-yl)-10,10-dimethyl-1²-(2-(4-methylpiperazin-1-yl)pyridin-4-yl)-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(110 mg, 0.14 mmol) and TFA (5.0 mL, 67.3 mmol) in DCM (5 mL) wasstirred at rt for 1 h, then concentrated under reduced pressure to give2-((6³S,4S,Z)-4-amino-10,10-dimethyl-1²-(2-(4-methylpiperazin-1-yl)pyridin-4-yl)-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-1¹-yl)-2-methylpropanenitrile(96 mg, 100% yield) as an oil. LCMS (ESI): m/z [M+H] calc'd forC₃₈H₄₇N₉O₃S. 709.4; found 710.5.

Step 11

A mixture of2-((6³S,4S,Z)-4-amino-10,10-dimethyl-1²-(2-(4-methylpiperazin-1-yl)pyridin-4-yl)-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-1¹-yl)-2-methylpropanenitrile(110 mg, 0.16 mmol), (1S,2S)-2-methylcyclopropane-1-carboxylic acid (47mg, 0.47 mmol), COMU (66 mg, 0.16 mmol), DIPEA (1.00 g, 7.75 mmol) inDMF (5 mL) was stirred at rt for 2 h. H₂O (10 mL) was added and themixture was extracted with EtOAc (2×10 mL). The combined organic layerswere washed with brine (2×10 mL), dried over Na₂SO₄ and filtered. Thefiltrate was concentrated under reduced pressure and the residue waspurified by preparative-HPLC to give(1S,2S)—N-((6³S,4S,Z)-1¹-(2-cyanopropan-2-yl)-10,10-dimethyl-1²-(2-(4-methylpiperazin-1-yl)pyridin-4-yl)-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2-methylcyclopropane-1-carboxamide(21 mg, 17% yield) as a solid. LCMS (ESI): m/z [M+H] calc'd forC₄₃H₅₃N₉O₄S. 791.4; found 792.4; ¹H-NMR (400 MHz, DMSO-d₆) δ 8.51 (d,J=7.1 Hz, 2H), 8.21 (d, J=5.0 Hz, 1H), 8.16 (d, J=5.0 Hz, 1H), 7.89 (d,J=1.8 Hz, 1H), 7.83 (d, J=1.5 Hz, 2H), 7.06 (s, 1H), 6.96 (s, 1H),6.74-6.68 (m, 1H), 5.54 (q, J=8.6 Hz, 1H), 5.04 (d, J=12.2 Hz, 1H), 4.20(q, J=12.4 Hz, 2H), 3.66 (dd, J=16.0, 10.9 Hz, 1H), 3.55 (s, 5H), 3.50(d, J=10.9 Hz, 1H), 3.35 (s, 1H), 3.15 (dd, J=14.8, 9.2 Hz, 1H), 2.93(dd, J=14.4, 6.3 Hz, 1H), 2.77 (s, 1H), 2.38 (dd, J=10.9, 5.5 Hz, 4H),2.20 (d, J=5.3 Hz, 3H), 2.09 (s, 1H), 2.06 (s, 1H), 2.09-1.99 (m, 3H),1.82-1.51 (d, J=4.0 Hz, 4H), 1.07 (s, 4H), 0.90-0.86 (d, J=3.0 Hz, 4H),0.55 (d, J=6.8 Hz, 1H), 0.47-0.38 (m, 3H).

Examples A221 and A222. Synthesis of(1S,2S)-2-(difluoromethyl)-N-((6³S,4S,Z)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)cyclopropane-1-carboxamideand(1R,2R)-2-(difluoromethyl)-N-((6³S,4S,Z)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)cyclopropane-1-carboxamide

Step 1

To a mixture of(6³S,4S,Z)-4-amino-1′-ethyl-1²-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-5,7-dione (200 mg, 0.27 mmol) andtrans-2-(difluoromethyl)cyclopropane-1-carboxylic acid (56 mg, 0.41mmol) in DMF (8 mL) at 0° C. was added DIPEA (177 mg, 1.37 mmol)dropwise, followed by COMU (235 mg, 0.55 mmol). The mixture was allowedto warm to rt and stirred for 1 h, then diluted with EtOAc (10 mL) andH₂O (50 mL). The aqueous and organic layers were separated and theaqueous layer was extracted with EtOAc (3×10 mL). The combined organiclayers were washed with brine (2×10 mL), dried over anhydrous Na₂SO₄ andfiltered. The filtrate was concentrated under reduced pressure and theresidue was purified by preparative-HPLC to give(1S,2S)-2-(difluoromethyl)-N-((6³S,4S,Z)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)cyclopropane-1-carboxamide(32 mg, 27% yield) and(1R,2R)-2-(difluoromethyl)-N-((6³S,4S,Z)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)cyclopropane-1-carboxamide (31 mg, 27%yield) both as solids. LCMS (ESI): m/z [M+H] calc'd for C₄₄H₅₆F₂N₈O₅S.846.4; found 847.3; ¹H NMR (400 MHz, DMSO-d₆) δ 8.80 (s, 1H), 8.46 (dd,J=13.4, 2.2 Hz, 2H), 7.83 (s, 2H), 7.73 (dd, J=8.7, 1.6 Hz, 1H), 7.56(d, J=8.6 Hz, 1H), 7.23 (d, J=2.8 Hz, 1H), 5.95 (d, J=5.3 Hz, 1H), 5.57(t, J=9.1 Hz, 1H), 5.12 (d, J=12.2 Hz, 1H), 4.14 (d, J=6.4 Hz, 5H), 3.57(s, 2H), 3.21 (s, 4H), 2.93 (d, J=14.3 Hz, 2H), 2.78-2.68 (m, 1H), 2.67(p, 0=1.9 Hz, 3H), 2.46-2.28 (m, 3H), 2.25 (s, 2H), 2.17-1.92 (m, 2H),1.79 (s, 2H), 1.66 (dt, J=9.7, 5.0 Hz, 1H), 1.51 (t, J=9.0 Hz, 1H), 1.33(d, J=6.1 Hz, 4H), 1.24 (d, J=5.6 Hz, 2H), 0.96 (s, 1H), 0.95-0.72 (m,6H), 0.35 (s, 3H) and LCMS (ESI): m/z [M+H] calc'd for C₄₄H₅₆F₂N₈O₅S.846.4; found 847.3; ¹H NMR (400 MHz, DMSO-d₆) δ 8.87 (d, J=9.0 Hz, 1H),8.46 (dd, J=11.3, 2.2 Hz, 2H), 7.80 (s, 2H), 7.73 (dd, 0=8.7, 1.6 Hz,1H), 7.56 (d, J=8.7 Hz, 1H), 7.23 (d, J=2.9 Hz, 1H), 6.09 (d, J=5.1 Hz,1H), 5.94 (d, 0=5.1 Hz, 1H), 5.80 (d, J=5.2 Hz, 1H), 5.59 (t, J=9.1 Hz,1H), 5.13 (d, J=12.2 Hz, 1H), 4.50-4.06 (m, 5H), 3.57 (s, 2H), 3.21 (s,4H), 2.93 (d, J=14.1 Hz, 2H), 2.67 (p, J=1.9 Hz, 4H), 2.39-2.15 (m, 4H),2.17-1.82 (m, 2H), 1.77 (d, J=16.1 Hz, 2H), 1.44 (d, J=43.3 Hz, 1H),1.35-1.24 (m, 4H), 1.15-0.91 (m, 2H), 0.91 (s, 6H), 0.34 (s, 3H).

Example A173. Synthesis of(2S)—N-((6³S,4S,Z)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2,4-dimethylpiperazine-1-carboxamide

Step 1

To a mixture of(6³S,4S,Z)-4-amino-1′-ethyl-1²-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-5,7-dione (80 mg, 0.11 mmol) in DCM (6 mL) at0° C. was added pyridine (2 mL), then 4-nitrophenyl carbonochloridate(55 mg, 0.28 mmol) in portions. The mixture was allowed to warm to rtand stirred for 1 h at room temperature, then washed with 1M NaHSO₄ (10mL) and H₂O (10 mL). The organic layer was concentrated under reducedpressure, to give 4-nitrophenyl((6³S,4S,Z)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(98 mg, crude) as a solid. LCMS (ESI): m/z [M+H] calc'd for C₄₆H₅₅N₉O₈S.893.4; found 894.2.

Step 2

To a mixture of 4-nitrophenyl((6³S,4S,Z)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(98 mg, 0.11 mmol) and (S)-1,3-dimethylpiperazine (63 mg, 0.55 mmol) inACN (5 mL) at 0° C. was added DIPEA (43 mg, 0.33 mmol) in ACN (2 mL. Thecrude product was purified by preparative-HPLC to give(2S)—N-((6³S,4S,Z)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2,4-dimethylpiperazine-1-carboxamide(13 mg, 13% yield) as a solid. LCMS (ESI): m/z [M+H] calc'd forC₄₆H₆₄N₁₀O₅S. 868.5; found 869.3; ¹H NMR (400 MHz, DMSO-d₆) δ 8.47 (s,2H), 7.80 (s, 1H), 7.73 (s, 1H), 7.55 (s, 1H), 7.23 (s, 1H), 6.81-6.70(m, 1H), 5.33-5.25 (m, 1H), 4.99 (s, 1H), 4.40-3.97 (m, 6H), 3.72 (s,1H), 3.61-3.47 (m, 3H), 3.31-3.22 (m, 8H), 3.02-2.72 (m, 5H), 2.66 (s,1H), 2.60-2.51 (m, 3H), 2.49-2.37 (m, 2H), 2.35-2.13 (m, 6H), 2.12-1.97(m, 2H), 1.95-1.66 (m, 3H), 1.55 (s, 1H), 1.33 (s, 3H), 1.27-1.19 (m,4H), 0.99-0.82 (m, 6H), 0.33 (s, 3H).

Example A225. Synthesis of(1S,2S)—N-((6³S,3S,4S,Z)-3-ethoxy-1′-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2-methylcyclopropane-1-carboxamide

Step 1

To a mixture of ethyl(2S,3S)-2-[bis[(4-methoxyphenyl)methyl]amino]-3-(4-bromo-1,3-thiazol-2-yl)-3-hydroxypropanoate(1.00 g, 1.9 mmol) and Ag₂O (4.33 g, 18.7 mmol) in ACN (10 mL) at 0° C.under an atmosphere of N₂ was added ethyl iodide (2.91 g, 18.7 mmol)dropwise. The mixture was heated to 80° C. and stirred for 4 h, thenfiltered and the filter cake was washed with ACN (3×5 mL). The filtratewas concentrated under reduced pressure and the residue was purified bysilica gel column chromatography to give ethyl(2S,3S)-2-[bis[(4-methoxyphenyl)methyl]amino]-3-(4-bromo-1,3-thiazol-2-yl)-3-ethoxypropanoate(557 mg, 53% yield) as an oil. LCMS (ESI): m/z [M+H] calc'd forC₂₆H₃₁BrN₂O₅S. 562.1; found 563.2.

Step 2

Into a 40 mL sealed tube were added ethyl(2S,3S)-2-[bis[(4-methoxyphenyl)methyl]amino]-3-(4-bromo-1,3-thiazol-2-yl)-3-ethoxypropanoate(530 mg) and TFA (10 mL) under an atmosphere of N₂. The mixture washeated to 80° C. and stirred overnight, then concentrated under reducedpressure to give ethyl(2S,3S)-2-amino-3-(4-bromo-1,3-thiazol-2-yl)-3-ethoxypropanoate that wasused directly in the next step without further purification. LCMS (ESI):m/z [M+H] calc'd for C₁₀H₁₅BrN₂O₃S. 322.0; found 323.0.

Step 3

A mixture of ethyl(2S,3S)-2-amino-3-(4-bromo-1,3-thiazol-2-yl)-3-ethoxypropanoate (890 mg,2.8 mmol), LiOH.H₂O (1.16 g, 27.6 mmol), MeOH (6 mL), THF (2 mL) and H₂O(2 mL) was stirred at 45° C. for 2 h. The mixture was concentrated underreduced pressure to give(2S,3S)-2-amino-3-(4-bromo-1,3-thiazol-2-yl)-3-ethoxypropanoic acid(that was used directly in the next step without further purification.LCMS (ESI): m/z [M+H] calc'd for C₈H₁₁BrN₂O₃S. 294.0; found 294.9.

Step 4

To a mixture of(2S,3S)-2-amino-3-(4-bromo-1,3-thiazol-2-yl)-3-ethoxypropanoic acid (890mg, 3.0 mmol), NaHCO₃ (507 mg, 6.0 mmol) and DMAP (37 mg, 0.3 mmol) inTHF/H₂O (1:1) at 0° C. was added (Boc)₂O (1.97 g, 9.0 mmol). The mixturewas warmed to rt and stirred overnight then concentrated under reducedpressure to remove THF and the residue was acidified to pH ˜6 with HCl.The mixture was extracted with EtOAc (3×5 mL) and the combined organiclayers were dried over anhydrous Na₂SO₄ and filtered. The filtrate wasconcentrated under reduced pressure and the residue was purified bysilica gel column chromatography to give(2S,3S)-3-(4-bromo-1,3-thiazol-2-yl)-2-[(tert-butoxycarbonyl)amino]-3-ethoxypropanoicacid (369 mg, 31% yield) as a solid. LCMS (ESI): m/z [M+H] calc'd forC₁₃H₁₉BrN₂O₅S 394.0; found 395.0.

Step 5

To a mixture of3-(1-ethyl-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)-2,2-dimethylpropyl-(S)-hexahydropyridazine-3-carboxylate(584 mg, 0.91 mmol) and(2S,3S)-3-(4-bromo-1,3-thiazol-2-yl)-2-[(tert-butoxycarbonyl)amino]-3-ethoxypropanoicacid (360 mg, 0.91 mmol) in DMF at 0° C. was added DIPEA (1.59 mL, 9.1mmol) and HATU (693 mg, 1.8 mmol). The mixture was allowed to warm to rtand stirred for 1 h at room temperature, then cooled to 0° C. and H2Oadded. The mixture was extracted with EtOAc (2×5 mL) and the combinedorganic layers were washed with brine (3×10 mL), dried over anhydrousNa₂SO₄ and filtered. The filtrate was concentrated under reducedpressure and the residue was purified by silica gel columnchromatography to give3-(1-ethyl-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)-2,2-dimethylpropyl(S)-1-((2S,3S)-3-(4-bromothiazol-2-yl)-2-((tert-butoxycarbonyl)amino)-3-ethoxypropanoyl)hexahydropyridazine-3-carboxylate(410 mg, 46%) yield as a solid. LCMS (ESI): m/z [M+H] calc'd forC₄₇H₆₆BBrN₆O₉S. 980.4; found 981.3.

Step 6

Into a 50 mL Schlenk tube were added3-(1-ethyl-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)-2,2-dimethylpropyl(S)-1-((2S,3S)-3-(4-bromothiazol-2-yl)-2-((tert-butoxycarbonyl)amino)-3-ethoxypropanoyl)hexahydropyridazine-3-carboxylate(390 mg, 0.4 mmol), Pd(DTBpf)Cl₂ (78 mg, 0.12 mmol), K₃PO₄ (211 mg, 1.0mmol), toluene (9 mL), 1,4-dioxane (3 mL) and H₂O (3 mL) under anatmosphere of Ar. The mixture was heated to 60° C. and stirred for 1 h,then extracted with EtOAc (3×15 mL). The combined organic layers weredried over anhydrous Na2SO4, filtered, the filtrate was concentratedunder reduced pressure and the residue was purified by silica gel columnchromatography to give tert-butyl((6³S,3S,4S,Z)-3-ethoxy-1′-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(96 mg, 31% yield) as a solid. LCMS (ESI): m/z [M+H] calc'd forC₄₁H₅₄N₆O₇S. 774.4; found 775.4.

Step 7

A mixture of tert-butyl((6³S,3S,4S,Z)-3-ethoxy-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate (92 mg, 0.12 mmol), HCl in1,4-dioxane (2.5 mL) and 1,4-dioxane (2.5 mL) was stirred at rt for 1 h.The mixture was concentrated under reduced pressure to give tert-butyl((6³S,3S,4S,Z)-3-ethoxy-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(100 mg), which was used directly in the next step without furtherpurification. LCMS (ESI): m/z [M+H] calc'd for C₃₆H₄₆N₆O₅S. 674.3; found675.3.

Step 8

To a mixture of tert-butyl((6³S,3S,4S,Z)-3-ethoxy-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate (90 mg, 0.13 mmol) and(1S,2S)-2-methylcyclopropane-1-carboxylic acid (27 mg, 0.27 mmol) in DMFat 0° C. was added DIPEA (172 mg, 1.3 mmol) and HATU (101 mg, 0.27mmol). The mixture was allowed to warm to rt and stirred for 1 h, thencooled to 0° C., H2O added and the mixture extracted with EtOAc (2×5mL). The combined organic layers were washed with brine (3×20 mL), driedover anhydrous Na₂SO₄ and filtered. The filtrate was concentrated underreduced pressure and the residue was purified by preparative-HPLC togive(1S,2S)—N-((6³S,3S,4S,Z)-3-ethoxy-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2-methylcyclopropane-1-carboxamide(21 mg, 21% yield) as a solid. LCMS (ESI): m/z [M+H] calc'd forC₄₁H₅₂N₆O₆S. 756.4; found 757.6; ¹H NMR (400 MHz, DMSO-d₆) δ 8.76 (dd,J=4.8, 1.7 Hz, 1H), 8.50 (s, 1H), 7.91 (s, 1H), 7.84-7.71 (m, 3H),7.61-7.56 (m, 1H), 7.55-7.50 (m, 1H), 5.91-5.85 (m, 1H), 5.20-5.14 (m,1H), 4.92 (s, 1H), 4.36-4.21 (m, 3H), 4.17-4.07 (m, 2H), 3.68-3.56 (m3H), 3.54-3.46 (m, 1H), 3.22 (s, 3H), 2.90-2.73 (m, 2H), 2.09-2.03 (m,1H), 1.88-1.73 (m, 3H), 1.55-1.43 (m, 1H), 1.37 (d, J=6.0 Hz, 3H), 1.20(t, J=6.9 Hz, 3H), 1.04 (s, 4H), 0.93-0.71 (m, 7H), 0.51 (d, J=6.3 Hz,1H), 0.38 (s, 3H).

Example A227. Synthesis of(1R,2R,3S)—N-((6³S,4S,Z)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2,3-dimethylcyclopropane-1-carboxamide

Step 1

To a mixture of(6³S,4S,Z)-4-amino-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-5,7-dione(126 mg, 0.20 mmol) and (1R,2R,3S)-2,3-dimethylcyclopropane-1-carboxylicacid (34 mg, 0.30 mmol) in DMF (5 mL) at 0° C. was added DIPEA (129 mg,1.0 mmol) and COMU (171 mg, 0.4 mmol). The mixture was warmed to rt andstirred for 1 h, then diluted with EtOAc (20 mL) and H₂O (20 mL). Theaqueous layer was extracted with EtOAc (3×10 mL) and the combinedorganic layers were washed with brine (2×10 mL), dried over anhydrousNa₂SO₄ and filtered. The filtrate was concentrated under reducedpressure and the residue was purified by preparative-HPLC to give(1R,2R,3S)—N-((6³S,4S,Z)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2,3-dimethylcyclopropane-1-carboxamide(29 mg, 25% yield) as a solid. LCMS (ESI): m/z [M+H] calc'd forC₄₀H₅₀N₆O₅S. 726.4; found 727.3; ¹H NMR (400 MHz, DMSO-d₆) δ 8.75 (dd,J=4.8, 1.7 Hz, 1H), 8.49 (d, J=1.6 Hz, 1H), 8.40 (d, J=9.0 Hz, 1H), 7.82(s, 3H), 7.58 (d, J=8.6 Hz, 1H), 7.52 (dd, J=7.7, 4.7 Hz, 1H), 5.56 (t,J=9.0 Hz, 1H), 5.07 (d, J=12.2 Hz, 1H), 4.44-3.99 (m, 5H), 3.57 (s, 1H),3.25 (s, 1H), 3.16 (d, J=9.3 Hz, 3H), 2.94 (d, J=14.3 Hz, 1H), 2.81-2.70(m, 1H), 2.67 (p, J=1.9 Hz, 1H),2.44-2.27 (m, 1H), 2.16-2.01 (m, 1H),1.78 (s, 2H), 1.53 (s, 1H), 1.37 (d, J=6.0 Hz, 3H), 1.26-1.13 (m, 3H),1.07 (dd, J=9.4, 5.4 Hz, 6H), 0.93-0.77 (m, 6H), 0.32 (s, 3H).

Example A372. Synthesis of(1r,2R,3S)—N-((2²R,6³S,4S)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-2²-(methoxymethyl)-10,10-dimethyl-5,7-dioxo-2¹,2²,2³,2⁶,6¹,6²,6³,6⁴,6⁵,6⁶-decahydro-1¹H-8-oxa-1(5,3)-indola-6(1,3)-pyridazina-2(5,1)-pyridinacycloundecaphane-4-yl)-2,3-dimethylcyclopropane-1-carboxamide

Step 1.

To a mixture of methyl (R)-2-aminopent-4-enoate (8.0 g, 36.5 mmol) and(Z)-(2,3-dibromoprop-1-en-1-yl)benzene (15.1 g, 54.8 mmol) in MECN (80mL) under an atmosphere of N₂ was added Cs₂CO₃ (35.7 g, 109.6 mmol) andKI (12.13 g, 73.1 mmol) in portions. The mixture was heated to 80° C.and stirred overnight, then filtered and the filter cake was washed withMeCN (3×20 mL). The filtrate was concentrated under reduced pressure andthe residue was purified by silica gel column chromatography to givemethyl (R,Z)-2-((2-bromo-3-phenylallyl)amino)pent-4-enoate (12.0 g, 91%yield) as an oil. LCMS (ESI): m/z [M+H]⁺ calc'd for C₁₅H₁₈BrNO₂ 323.1;found 324.1.

Step 2.

To a mixture of methyl(R,Z)-2-((2-bromo-3-phenylallyl)amino)pent-4-enoate (12.0 g, 37.0 mmol)and BnBr (12.66 g, 74.0 mmol) in MeCN (120 mL) under an atmosphere of N₂was added Cs₂CO₃ (24.12 g, 74.0 mmol) and KI (6.14 g, 37.0 mmol) inportions. The mixture was stirred at room temperature overnight, thenfiltered and the filter cake was washed with MeCN (3×20 mL). Thefiltrate was concentrated under reduced pressure and the residue waspurified by silica gel column chromatography to give methyl(R,Z)-2-(benzyl(2-bromo-3-phenylallyl)amino)pent-4-enoate (6.0 g, 35%yield) as an oil. LCMS (ESI): m/z [M+H]⁺ calc'd for C₂₂H₂₄BrNO₂ 415.1;found 416.1 [for ⁸¹Br].

Step 3.

To a mixture of methyl(R,Z)-2-(benzyl(2-bromo-3-phenylallyl)amino)pent-4-enoate (5.8 g, 14.0mmol) and[1,3-bis(2,4,6-trimethylphenyl)imidazolidin-2-ylidene]dichloro[[2-(propan-2-yloxy)phenyl]methylidene]ruthenium(2.63 g, 4.2 mmol) in toluene (580 mL) under an atmosphere of Ar washeated to 60° C. and stirred for 30 min. The mixture was concentratedunder reduced pressure and the residue was purified by preparative-HPLCto give methyl(R)-1-benzyl-5-bromo-1,2,3,6-tetrahydropyridine-2-carboxylate (3.7 g,77% yield) as an oil. LCMS (ESI): m/z [M+H]⁺ calc'd for C₁₄H₁₆BrNO₂309.0; found 310.1.

Step 4.

To a mixture of methyl(R)-1-benzyl-5-bromo-1,2,3,6-tetrahydropyridine-2-carboxylate (3.7 g,11.9 mmol) and CaCl₂ (2.65 g, 23.9 mmol) in EtOH (22 mL) and THF (15 mL)at 0° C. under an atmosphere of N₂ was added NaBH₄ (1.80 g, 47.7 mmol)in portions. The mixture was warmed to room temperature and stirred for2 h, then cooled to 0° C., and MeOH and H₂O were added. The mixture wasextracted with DCM (2×50 mL), dried over anhydrous Na₂SO₄ and filtered.The filtrate was concentrated under reduced pressure and the residue waspurified by preparative HPLC to give(R)-(1-benzyl-5-bromo-1,2,3,6-tetrahydropyridin-2-yl)methanol (2.8 g,75% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd for C₁₃H₁₆BrNO281.0; found 282.3.

Step 5.

To a mixture of(R)-(1-benzyl-5-bromo-1,2,3,6-tetrahydropyridin-2-yl)methanol (1.0 g,3.5 mmol) in THF (10 mL) at 0° C. was added NaH, 60% dispersion in oil(0.26 g, 10.6 mmol). The mixture was stirred for 15 min, then Mel (0.75g, 5.3 mmol) was added and the mixture was allowed to warm to roomtemperature and stirred for 1 h. The mixture was cooled to 0° C.,saturated NH₄Cl was added and the mixture was extracted with EtOAc (2×20mL). The combined organic layers were dried over anhydrous Na₂SO₄,filtered, the filtrate was concentrated under reduced pressure and theresidue was purified by preparative-HPLC to give(R)-1-benzyl-5-bromo-2-(methoxymethyl)-1,2,3,6-tetrahydropyridine (1.0g, 86% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd for C₁₄H₁₈BrNO295.1; found 296.2.

Step 6.

A mixture of(R)-1-benzyl-5-bromo-2-(methoxymethyl)-1,2,3,6-tetrahydropyridine (1.0g, 3.4 mmol) and 1-chloroethyl chloroformate (2 mL, 14.0 mmol) in DCM(10 mL) under an atmosphere of N₂ was stirred at room temperatureovernight. The mixture was washed with brine (2×10 mL) and the combinedaqueous layers were extracted with DCM (10 mL). The combined organiclayers were dried over anhydrous Na₂SO₄, filtered and the filtrate wasconcentrated under reduced pressure. The residue was dissolved in MeOH(10 mL), the mixture was heated to 50° C. under an atmosphere of N₂ andstirred for 2 h, then concentrated under reduced pressure and theresidue was purified by preparative-HPLC to give(R)-5-bromo-2-(methoxymethyl)-1,2,3,6-tetrahydropyridine (500 mg, 72%yield) as an oil. LCMS (ESI): m/z [M+H]⁺ calc'd for C₇H₁₂BrNO 205.0;found 206.1.

Step 7.

To a mixture of (R)-5-bromo-2-(methoxymethyl)-1,2,3,6-tetrahydropyridine(600 mg, 2.9 mmol) and tert-butyl N-[(3S)-2-oxooxetan-3-yl]carbamate(382 mg, 2.0 mmol) in MeCN (6 mL) under an atmosphere of N₂ was stirredovernight. The mixture was concentrated under reduced pressure and theresidue was purified by preparative-HPLC to give(S)-3-((R)-5-bromo-2-(methoxymethyl)-3,6-dihydropyridin-1(2H)-yl)-2-((tert-butoxycarbonyl)amino)propanoicacid (500 mg, 39% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₁₅H₂₅BrN₂O₅ 392.1; found 393.1.

Step 8.

To a mixture of3-(1-ethyl-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)-2,2-dimethylpropyl(S)-hexahydropyridazine-3-carboxylate (1.25 g, 2.07 mmol) and(S)-3-((R)-5-bromo-2-(methoxymethyl)-3,6-dihydropyridin-1(2H)-yl)-2-((tert-butoxycarbonyl)amino)propanoic acid (0.49 g, 1.24mmol) and DIPEA (2.67 g, 20.7 mmol) in DMF (15 mL) under an atmosphereof N₂ was added HATU (0.79 g, 2.07 mmol) in portions. The mixture waswarmed to room temperature and stirred for 1 h, then H₂O added and themixture was extracted with EtOAc (50 mL). The organic layer was washedwith brine (3×50 mL), dried over anhydrous Na₂SO₄ and filtered. Thefiltrate was concentrated under reduced pressure and the residue waspurified by silica gel column chromatography to give3-(1-ethyl-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)-2,2-dimethylpropyl(S)-1-((S)-3-((R)-5-bromo-2-(methoxymethyl)-3,6-dihydropyridin-1(2H)-yl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylate(880 mg, 39% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₄₉H₇₂BBrN₆O₉ 980.5; found 981.4 [for ⁸¹Br].

Step 9.

To a mixture of3-(1-ethyl-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)-2,2-dimethylpropyl(S)-1-((S)-3-((R)-5-bromo-2-(methoxymethyl)-3,6-dihydropyridin-1(2H)-yl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylate(850 mg, 0.87 mmol) and K₃PO₄ (460 mg, 2.17 mmol) in toluene (4.5 mL),1,4-dioxane (1.5 mL) and H₂O (1.5 mL) under an atmosphere of N₂ wasadded Pd(dppf)Cl₂ (64 mg, 0.09 mmol) in portions. The mixture was heatedto 70° C. and stirred for 3 h at 70° C., then concentrated under reducedpressure and the residue was purified by silica gel columnchromatography to give tert-butyl((2²R,6³S,4S)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-2²-(methoxymethyl)-10,10-dimethyl-5,7-dioxo-2¹,2²,2³,2⁶,6¹,6²,6³,6⁴,6⁵,6⁶-decahydro-1¹H-8-oxa-1(5,3)-indola-6(1,3)-pyridazina-2(5,1)-pyridinacycloundecaphane-4-yl)carbamate(220 mg, 30% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₄₃H₆₀N₆O₇ 772.5; found 773.4.

Step 10.

A mixture of tert-butyl((2²R,6³S,4S)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-2²-(methoxymethyl)-10,10-dimethyl-5,7-dioxo-2¹,2²,2³,2⁶,6¹,6²,6³,6⁴,6⁵,6⁶-decahydro-1¹H-8-oxa-1(5,3)-indola-6(1,3)-pyridazina-2(5,1)-pyridinacycloundecaphane-4-yl)carbamate(100 mg, 0.13 mmol) and HCl in 1,4-dioxane (2 mL) in DCM (2 mL) wasstirred at room temperature for 1 h. The mixture was concentrated underreduced pressure to give(2²R,6³S,4S)-4-amino-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-2²-(methoxymethyl)-10,10-dimethyl-2¹,2²,2³,2⁶,6¹,6²,6³,6⁴,6⁵,6⁶-decahydro-1¹H-8-oxa-1(5,3)-indola-6(1,3)-pyridazina-2(5,1)-pyridinacycloundecaphane-5,7-dione(119 mg) as a solid, that was used directly in the next step withoutfurther purification.

Step 11.

To a mixture of(2²R,6³S,4S)-4-amino-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-2²-(methoxymethyl)-10,10-dimethyl-2¹,2²,2³,2⁶,6¹,6²,6³,6⁴,6⁵,6⁶-decahydro-1¹H-8-oxa-1(5,3)-indola-6(1,3)-pyridazina-2(5,1)-pyridinacycloundecaphane-5,7-dione (20 mg, 0.18 mmol),(1r,2R,3S)-2,3-dimethylcyclopropanecarboxylic acid, and DIPEA (114 mg,0.89 mmol) in DMF (2 mL) was added HATU (81 mg, 0.21 mmol) in portions.The mixture was stirred at room temperature for 1 h, then purified bypreparative-HPLC to give(1r,2R,3S)—N-((2²R,6³S,4S)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-2²-(methoxymethyl)-10,10-dimethyl-5,7-dioxo-2¹,2²,2³,2⁶,6¹,6²,6³,6⁴,6⁵,6⁶-decahydro-1¹H-8-oxa-1(5,3)-indola-6(1,3)-pyridazina-2(5,1)-pyridinacycloundecaphane-4-yl)-2,3-dimethylcyclopropane-1-carboxamide(24 mg, 18% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₄₄H₆₀N₆O₆ 768.5; found 769.2; ¹H NMR (400 MHz, DMSO-d₆) δ 8.90-8.65 (m,1H), 8.10-7.95 (m, 1H), 7.90-7.75 (m, 1H), 7.57-7.45 (m, 3H), 7.40 (s,1H), 6.26 (s, 1H), 5.65-5.50 (m, 1H), 5.45-5.30 (m, 1H), 4.39-4.25 (m,1H), 4.24-4.20 (m, 1H), 4.19-4.10 (m, 1H), 4.09-4.00 (m, 1H), 3.88-3.76(m, 1H), 3.73-3.62 (m, 2H), 3.62-3.55 (m, 2H), 3.26 (s, 5H), 3.14-3.00(m, 2H), 2.96-2.87 (m, 1H), 2.79-2.70 (m, 5H), 2.43 (s, 1H), 2.15-2.06(m, 2H), 1.95-1.86 (m, 1H), 1.86-1.72 (m, 1H), 1.58-1.49 (m, 2H),1.40-1.30 (m, 3H), 1.10-1.00 (m, 6H), 0.99-0.90 (m, 6H), 0.82 (s, 3H),0.52 (s, 3H).

Example A373. Synthesis of(1r,2R,3S)—N-((63S,4S,Z)-1²-(5-(1-(2-(dimethylamino)ethyl)-4-hydroxypiperidin-4-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-1¹-ethyl-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2,3-dimethylcyclopropane-1-carboxamide

Step 1.

To a mixture of (S)-3-bromo-5-iodo-2-(1-methoxyethyl)pyridine (5.0 g,14.6 mmol) in THF (40 mL) at −78° C. under an atmosphere of N₂ was addedn-BuLi in hexanes (5.85 mL, 14.6 mmol) dropwise. The mixture was stirredat −78° C. for 1 h, then benzyl 4-oxopiperidine-1-carboxylate (6.82 g,29.2 mmol) was added. The mixture was allowed to warm to 0° C. SaturatedNH₄Cl (3 mL) was added, the mixture was diluted with H₂O (20 mL) andextracted with EtOAc (3×20 mL). The combined organic layers were driedover anhydrous Na₂SO₄, filtered, the filtrate was concentrated underreduced pressure and the residue was purified by silica gel columnchromatography to give benzyl(S)-4-(5-bromo-6-(1-methoxyethyl)pyridin-3-yl)-4-hydroxypiperidine-1-carboxylate(800 mg, 12% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₂₁H₂₅BrN₂O₄ 448.1; found 449.2.

Step 2.

To a mixture of tert-butyl((6³S,4S,Z)-10,10-dimethyl-5,7-dioxo-1²-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(500 mg, 0.72 mmol) and benzyl(S)-4-(5-bromo-6-(1-methoxyethyl)pyridin-3-yl)-4-hydroxypiperidine-1-carboxylate(648 mg, 1.4 mmol) in toluene (9 mL), H₂O (3 mL) and 1,4-dioxane (3 mL)under an atmosphere of N₂ was added K₃PO₄ (459 mg, 2.16 mmol) andPd(dppf)Cl₂.CH₂Cl₂ (59 mg, 0.07 mmol) in portions. The mixture washeated to 60° C. and stirred for 4 h, then diluted with H₂O (10 mL) andextracted with EtOAc (3×10 mL). The combined organic layers were driedover anhydrous Na₂SO₄, filtered, the filtrate was concentrated underreduced pressure and the residue was purified by silica gel columnchromatography to give benzyl4-(5-((6³S,4S,Z)-4-((tert-butoxycarbonyl)amino)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-1²-yl)-6-((S)-1-methoxyethyl)pyridin-3-yl)-4-hydroxypiperidine-1-carboxylate(1.16 g, 75% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₅₀H₆₁N₇O₉S. 935.4; found 936.4.

Step 3.

To a mixture of benzyl4-(5-((6³S,4S,Z)-4-((tert-butoxycarbonyl)amino)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-1²-yl)-6-((S)-1-methoxyethyl)pyridin-3-yl)-4-hydroxypiperidine-1-carboxylate(1.1 g, 1.18 mmol) and Cs₂CO₃ (1.91 g, 5.88 mmol) in DMF (15 mL) at 0°C. was added iodoethane (0.64 g, 4.1 mmol) dropwise. The mixture waswarmed to room temperature and stirred for 2 h, then diluted with H₂O(15 mL) and extracted with EtOAc (3×20 mL). The combined organic layerswere dried over anhydrous Na₂SO₄, filtered, the filtrate wasconcentrated under reduced pressure and the residue was purified bypreparative-TLC to give benzyl4-(5-((6³S,4S,Z)-4-((tert-butoxycarbonyl)amino)-1¹-ethyl-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-1²-yl)-6-((S)-1-methoxyethyl)pyridin-3-yl)-4-hydroxypiperidine-1-carboxylate(110 mg, 9% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₅₂H₆₅N₇O₉S. 963.5; found 964.4.

Step 4.

To a mixture of benzyl4-(5-((6³S,4S,Z)-4-((tert-butoxycarbonyl)amino)-1¹-ethyl-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-1²-yl)-6-((S)-1-methoxyethyl)pyridin-3-yl)-4-hydroxypiperidine-1-carboxylate(110 mg, 0.11 mmol) in DCM (3 mL) at 0° C. was added TFA (1 mL)dropwise. The mixture was warmed to room temperature and stirred for 1.5h, then concentrated under reduced pressure to give benzyl4-(5-((6³S,4S,Z)-4-amino-1¹-ethyl-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-1²-yl)-6-((S)-1-methoxyethyl)pyridin-3-yl)-4-hydroxypiperidine-1-carboxylate(120 mg) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd for C₄₇H₅₇N₇O₇S863.4; found 864.5.

Step 5.

To a mixture of benzyl4-(5-((6³S,4S,Z)-4-amino-1′-ethyl-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-1²-yl)-6-((S)-1-methoxyethyl)pyridin-3-yl)-4-hydroxypiperidine-1-carboxylate(120 mg, 0.14 mmol) and (1R,2R,3S)-2,3-dimethylcyclopropane-1-carboxylicacid (32 mg, 0.28 mmol) in DMF (4 mL) at 0° C. was added DIPEA (180 mg,1.39 mmol) and HATU (158 mg, 0.42 mmol) in portions. The mixture waswarmed to room temperature and stirred for 2 h, then diluted with H₂O(10 mL) and extracted with EtOAc (3×10 mL). The combined organic layerswere dried over anhydrous Na₂SO₄. After filtration, the filtrate wasconcentrated under reduced pressure. The residue was purified by reverseflash chromatography with the following conditions: column, C18 silicagel; mobile phase, MECN in water (0.05% TFA), 0% to 100% gradient in 300min; detector, UV 254 nm. To afford benzyl4-(5-((6³S,4S,Z)-4-((1r,2R,3S)-2,3-dimethylcyclopropane-1-carboxamido)-1¹-ethyl-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-1²-yl)-6-((S)-1-methoxyethyl)pyridin-3-yl)-4-hydroxypiperidine-1-carboxylate(120 mg, 89% yield) as a brown solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₅₃H₆₅N₇O₈S. 959.4; found 960.4.

Step 6.

A mixture of benzyl4-(5-((6³S,4S,Z)-4-((1r,2R,3S)-2,3-dimethylcyclopropane-1-carboxamido)-1¹-ethyl-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-1²-yl)-6-((S)-1-methoxyethyl)pyridin-3-yl)-4-hydroxypiperidine-1-carboxylate(120 mg, 0.13 mmol) and Pd(OH)₂, 30% on carbon (80 mg, 0.25 mmol) inEtOAc (2 mL) was stirred under an atmosphere of H2 overnight. Themixture was filtered, the filter cake was washed with MeOH (3×8 mL) andthe filtrate was concentrated under reduced pressure to give(1r,2R,3S)—N-((6³S,4S,Z)-1¹-ethyl-1²-(5-(4-hydroxypiperidin-4-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2,3-dimethylcyclopropane-1-carboxamide(45 mg) as an oil. LCMS (ESI): m/z [M+H]⁺ calc'd for C₄₅H₅₉N₇O₆S 825.4;found 826.4.

Step 7.

To a mixture of(1r,2R,3S)—N-((6³S,4S,Z)-1¹-ethyl-1²-(5-(4-hydroxypiperidin-4-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2,3-dimethylcyclopropane-1-carboxamide(50 mg, 0.06 mmol) and tert-butyl N-methyl-N-(2-oxoethyl)carbamate (52mg, 0.31 mmol) in MeOH (2 mL) at 0° C. was added ZnCl₂ (83 mg, 0.61mmol) in portions. The mixture was warmed to room temperature andstirred for 30 min, then cooled to 0° C. and NaBH₃CN (11 mg, 0.18 mmol)added in portions. The mixture was warmed to room temperature andstirred for 2 h, then the residue was purified by preparative-HPLC togive tert-butyl(2-(4-(5-((6³S,4S,Z)-4-((1r,2R,3S)-2,3-dimethylcyclopropane-1-carboxamido)-1¹-ethyl-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-1²-yl)-6-((S)-1-methoxyethyl)pyridin-3-yl)-4-hydroxypiperidin-1-yl)ethyl)(methyl)carbamate(30 mg, 50% yield) as a yellow oil. LCMS (ESI): m/z [M+H]⁺ calc'd forC₅₃H₇₄N₈O₈S. 982.5; found 983.6.

Step 8.

To a mixture of tert-butyl(2-(4-(5-((6³S,4S,Z)-4-((1r,2R,3S)-2,3-dimethylcyclopropane-1-carboxamido)-1¹-ethyl-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-1²-yl)-6-((S)-1-methoxyethyl)pyridin-3-yl)-4-hydroxypiperidin-1-yl)ethyl)(methyl)carbamate(30 mg, 0.03 mmol) in FA (1.5 mL) at 0° C. was added Et₃SiH (18 mg, 0.16mmol). The mixture was warmed to room temperature and stirred for 40min, then concentrated under reduced pressure to give(1r,2R,3S)—N-((63S,4S,Z)-1¹-ethyl-1²-(5-(4-hydroxy-1-(2-(methylamino)ethyl)piperidin-4-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2,3-dimethylcyclopropane-1-carboxamide(25 mg) as a solid, which was used directly in the next step withoutfurther purification. LCMS (ESI): m/z [M+H]⁺ calc'd for C₄₈H₆₆N₈O₆S.882.5; found 883.6.

Step 9.

To a mixture of(1r,2R,3S)—N-((63S,4S,Z)-11-ethyl-1²-(5-(4-hydroxy-1-(2-(methylamino)ethyl)piperidin-4-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2,3-dimethylcyclopropane-1-carboxamide(30 mg, 0.03 mmol) and formaldehyde (31 mg, 1.0 mmol) in MeOH (3 mL) at0° C. was added ZnCl₂ (46 mg, 0.34 mmol) in portions. The mixture waswarmed to room temperature and stirred for 30 min, then cooled to 0° C.and NaBH₃CN (6.4 mg, 0.10 mmol) was added in portions. The mixture waswarmed to room temperature and stirred for 2 h, then purified bypreparative-HPLC to give(1r,2R,3S)—N-((63S,4S,Z)-1²-(5-(1-(2-(dimethylamino)ethyl)-4-hydroxypiperidin-4-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-1¹-ethyl-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2,3-dimethylcyclopropane-1-carboxamide(0.7 mg, 2% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₄₉H₆₈N₈O₆S. 896.5; found 897.4; ¹H NMR (400 MHz, DMSO-d₆) δ 8.83 (d,J=2.3 Hz, 1H), 8.49 (d, J=1.6 Hz, 1H), 8.38 (d, J=9.1 Hz, 2H), 7.81 (s,1H), 7.78-7.68 (m, 2H), 7.57 (d, J=8.7 Hz, 1H), 5.57 (t, J=9.3 Hz, 1H),5.06 (d, J=12.1 Hz, 1H), 4.36-4.07 (m, 6H), 3.56 (s, 2H), 3.24 (s, 3H),3.18-3.11 (m, 1H), 2.93 (d, J=14.5 Hz, 1H), 2.75-2.60 (m, 2H), 2.43 (d,J=11.4 Hz, 4H), 2.37 (s, 2H), 2.15 (s, 7H), 2.02 (d, J=16.7 Hz, 4H),1.78 (s, 2H), 1.66 (d, J=12.6 Hz, 2H), 1.50 (s, 1H), 1.36 (d, J=6.1 Hz,3H), 1.24 (s, 1H), 1.16 (s, 2H), 1.15-0.93 (m, 6H), 0.92-0.63 (m, 6H),0.31 (s, 3H).

Example A387. Synthesis of(1r,2R,3S)—N-((6³S,3S,4S,Z)-3-(2,2-difluoroethoxy)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2,3-dimethylcyclopropane-1-carboxamide

Step 1.

To a mixture of ethyl(2S,3S)-2-(bis(4-methoxybenzyl)amino)-3-(4-bromothiazol-2-yl)-3-hydroxypropanoate(1.0 g, 1.9 mmol) in MeCN (10 mL) at room temperature under anatmosphere of N₂ was added Ag₂O (2.17 g, 9.4 mmol) and allyl iodide(1.57 g, 9.36 mmol). The resulting mixture was heated to 60° C. andstirred for 16 h, then filtered, and the filter cake was washed withEtOAc (3×20 mL). The filtrate was concentrated under reduced pressureand the residue was purified by silica gel column chromatography to giveethyl(2S,3S)-3-(allyloxy)-2-(bis(4-methoxybenzyl)amino)-3-(4-bromothiazol-2-yl)propanoate(1.0 g, 93% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₂₇H₃₁BrN₂O₅S. 574.1 & 576.1; found 575.1 & 577.1.

Step 2.

To a mixture of ethyl(2S,3S)-3-(allyloxy)-2-(bis(4-methoxybenzyl)amino)-3-(4-bromothiazol-2-yl)propanoate(1.0 g, 1.7 mmol) in 1,4-dioxane (10 mL) and H₂O (10 mL) at 0° C. wasadded 2,6-lutidine (0.37 g, 3.48 mmol) and K₂OsO₄.2H₂O (0.03 g, 0.09mmol). The mixture was stirred at 0° C. for 15 min then NaIO₄ (1.49 g,6.95 mmol) was added in portions. The mixture was warmed to roomtemperature and stirred for 2.5 h, then diluted with H₂O (50 mL) andextracted with EtOAc (3×50 mL). The combined organic layers were washedwith brine (2×50 mL), dried over anhydrous Na₂SO₄, filtered and thefiltrate was concentrated under reduced pressure to give ethyl(2S,3S)-2-(bis(4-methoxybenzyl)amino)-3-(4-bromothiazol-2-yl)-3-(2-oxoethoxy)propanoate(1.2 g) as an oil, which was used directly in the next step withoutfurther purification. LCMS (ESI): m/z [M+H]⁺ calc'd for C₂₆H₂₉BrN₂O₆S.576.1 & 578.1; found 577.4 & 579.4.

Step 3.

To a mixture of ethyl(2S,3S)-2-(bis(4-methoxybenzyl)amino)-3-(4-bromothiazol-2-yl)-3-(2-oxoethoxy)propanoate(1.2 g, 2.1 mmol) in DCM (20 mL) at −15° C. under an atmosphere of N₂was added DAST (0.37 g, 2.3 mmol) dropwise. The mixture was warmed toroom temperature and stirred for 1.5 h, then re-cooled to 0° C. andfurther DAST (0.37 g, 2.3 mmol) added dropwise. The mixture was warmedto room temperature and stirred for 1 h, then cooled to 0° C. andsaturated NH₄Cl (2 mL) added. The mixture was diluted with H₂O (20 mL)and extracted with DCM (3×20 mL). The combined organic layers werewashed with brine (2×50 mL), dried over anhydrous Na₂SO₄ and filtered.The filtrate was concentrated under reduced pressure and the residue waspurified by preparative-TLC to give ethyl(2S,3S)-2-(bis(4-methoxybenzyl)amino)-3-(4-bromothiazol-2-yl)-3-(2,2-difluoroethoxy)propanoate(270 mg, 22% yield) as an oil. LCMS (ESI): m/z [M+H]⁺ calc'd forC₂₆H₂₉BrF₂N₂O₅S. 598.1 & 600.1; found 599.1 & 601.1.

Step 4.

A mixture of ethyl(2S,3S)-2-(bis(4-methoxybenzyl)amino)-3-(4-bromothiazol-2-yl)-3-(2,2-difluoroethoxy)propanoate(240 mg, 0.40 mmol) in TFA (3 mL) was heated to 80° C. and stirred for 8h, then concentrated under reduced pressure to give ethyl(2S,3S)-2-amino-3-(4-bromothiazol-2-yl)-3-(2,2-difluoroethoxy)propanoate(245 mg) as an oil, that was used directly in the next step withoutfurther purification. LCMS (ESI): m/z [M+H]⁺ calc'd forC₁₀H₁₃BrF₂N₂CO₃S. 358.0 & 360.0; found 359.0 & 361.0.

Step 5.

To a mixture of ethyl(2S,3S)-2-amino-3-(4-bromothiazol-2-yl)-3-(2,2-difluoroethoxy)propanoate(230 mg, 0.64 mmol) and NaHCO₃ (108 mg, 1.29 mmol) in H₂O (0.9 mL) andTHF (3 mL) was added (Boc)₂O (147 mg, 0.67 mmol). The mixture wasstirred at room temperature overnight, then diluted with H₂O (10 mL) andextracted with EtOAc (3×10 mL). The combined organic layers were washedwith brine (3×20 mL), dried over anhydrous Na₂SO₄ and filtered. Thefiltrate was concentrated under reduced pressure and the residue waspurified by preparative-TLC to give ethyl(2S,3S)-3-(4-bromothiazol-2-yl)-2-((tert-butoxycarbonyl)amino)-3-(2,2-difluoroethoxy)propanoate(145 mg, 49% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₁₅H₂₁BrF₂N₂O₅S. 458.0 & 460.0; found 459.0 & 461.0.

Step 6.

To a mixture of ethyl(2S,3S)-3-(4-bromothiazol-2-yl)-2-((tert-butoxycarbonyl)amino)-3-(2,2-difluoroethoxy)propanoate(140 mg, 0.31 mmol) in THF (1.5 mL) at 0° C. was added LiOH.H₂O (64 mg,1.53 mmol) in H₂O (1.5 mL) dropwise. The mixture was warmed to roomtemperature and stirred for 1 h, then acidified to pH ˜5 with aqueousHCl, then extracted with EtOAc (3×10 mL). The combined organic layerswere washed with brine (3×20 mL), dried over anhydrous Na₂SO₄, filteredand the filtrate was concentrated under reduced pressure to give(2S,3S)-3-(4-bromothiazol-2-yl)-2-((tert-butoxycarbonyl)amino)-3-(2,2-difluoroethoxy)propanoicacid (100 mg) as an oil. LCMS (ESI): m/z [M+H]⁺ calc'd forC₁₃H₁₇BrF₂N₂O₅S. 430.0 & 432.0; found 431.0 & 433.0.

Step 7.

To a mixture of3-(1-ethyl-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)-2,2-dimethylpropyl-(S)-hexahydropyridazine-3-carboxylate(134 mg, 0.22 mmol) and NMM (335 mg, 3.32 mmol) in DCM (6 mL) at 0° C.under an atmosphere of N₂ was added(2S,3S)-3-(4-bromothiazol-2-yl)-2-((tert-butoxycarbonyl)amino)-3-(2,2-difluoroethoxy)propanoicacid (95 mg, 0.22 mmol) and HOBT (6 mg, 0.04 mmol) and EDCl (85 mg, 0.44mmol). The mixture was warmed to room temperature and stirred overnight,then diluted with H₂O (10 mL) and extracted with DCM (3×10 mL). Thecombined organic layers were washed with brine (3×20 mL), dried overanhydrous Na₂SO₄ and filtered. The filtrate was concentrated underreduced pressure and the residue was purified by preparative-TLC to give3-(1-ethyl-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)-2,2-dimethylpropyl-(S)-1-((2S,3S)-3-(4-bromothiazol-2-yl)-2-((tert-butoxycarbonyl)amino)-3-(2,2-difluoroethoxy)propanoyl)hexahydropyridazine-3-carboxylate(70 mg, 36% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₄₇H₆₄BBrF₂N₆O₉S. 1016.4 & 1018.4; found 1017.3 & 1019.4.

Step 8.

To a mixture of mixture of3-(1-ethyl-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)-2,2-dimethylpropyl-(S)-1-((2S,3S)-3-(4-bromothiazol-2-yl)-2-((tert-butoxycarbonyl)amino)-3-(2,2-difluoroethoxy)propanoyl)hexahydropyridazine-3-carboxylate(50 mg, 0.05 mmol) and K₃PO₄ (31 mg, 0.15 mmol) in toluene (3 mL),1,4-dioxane (1 mL) and H₂O (1 mL) under an atmosphere of N₂ was addedPd(DtBPF)Cl₂ (13 mg, 0.02 mmol) in portions. The mixture was heated to60° C. and stirred for 1 h, then was extracted with EtOAc (3×10 mL). Thecombined organic layers were washed with brine (3×20 mL), dried overanhydrous Na₂SO₄ and filtered. The filtrate was concentrated underreduced pressure and the residue was purified by preparative-TLC to givetert-butyl((6³S,3S,4S,Z)-3-(2,2-difluoroethoxy)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(27 mg, 59% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₄₁H₅₂F₂N₆O₇S. 810.4; found 811.4.

Step 9.

To a mixture tert-butyl((6³S,3S,4S,Z)-3-(2,2-difluoroethoxy)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(25 mg, 0.02 mmol) in DCM (1.5 mL) at 0° C. was added TFA (0.5 mL)dropwise. The mixture was warmed to room temperature and stirred for 1h, then concentrated under reduced pressure to give(6³S,3S,4S,Z)-4-amino-3-(2,2-difluoroethoxy)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-5,7-dione(25 mg) as a solid, which was used directly in the next step withoutfurther purification. LCMS (ESI): m/z [M+H]⁺ calc'd for C₃₆H₄₄F₂N₆O₅S.710.3; found 711.3.

Step 10.

To a mixture of(6³S,3S,4S,Z)-4-amino-3-(2,2-difluoroethoxy)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-5,7-dione(25 mg, 0.04 mmol) and (1r,2R,3S)-2,3-dimethylcyclopropane-1-carboxylicacid (6 mg, 0.05 mmol) in DMF (1 mL) at 0° C. under an atmosphere of N₂was added DIPEA (45 mg, 0.35 mmol) and HATU (27 mg, 0.07 mmol) inportions. The mixture was warmed to room temperature and stirred for 2h, then H₂O (10 mL) added and the mixture was extracted with EtOAc (3×10mL). The combined organic layers were washed with brine (3×20 mL), driedover anhydrous Na₂SO₄ and filtered. The filtrate was concentrated underreduced pressure and the residue was purified by preparative-HPLC togive(1r,2R,3S)—N-((6³S,3S,4S,Z)-3-(2,2-difluoroethoxy)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2,3-dimethylcyclopropane-1-carboxamide(3.3 mg, 12% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₄₂H₅₂F₂N₆O₆S. 806.4; found 807.2; ¹H NMR (400 MHz, DMSO-d₆) δ 8.95-8.70(m, 1H), 8.50 (s, 1H), 8.13 (s, 1H), 7.95 (s, 1H), 7.88-7.79 (m, 2H),7.76 (d, J=8.6 Hz, 1H), 7.59 (d, J=8.7 Hz, 1H), 7.54 (dd, J=7.7, 4.8 Hz,1H), 7.27-6.88 (m, 1H), 6.36-6.04 (m, 1H), 5.97 (d, J=9.9 Hz, 1H), 5.20(d, J=12.8 Hz, 2H), 4.25 (dd, J=20.3, 10.5 Hz, 4H), 4.14-3.92 (m, 2H),3.92-3.73 (m, 1H), 3.59 (q, J=10.9 Hz, 2H), 3.20 (s, 3H), 2.81 (d,J=13.7 Hz, 2H), 2.12-1.93 (m, 1H), 1.78 (d, J=23.7 Hz, 2H), 1.59-1.45(m, 2H), 1.38 (d, J=6.1 Hz, 3H), 1.21 (d, J=20.3 Hz, 1H), 1.16-1.01 (m,7H), 0.95-0.73 (m, 6H), 0.43 (s, 3H).

Example A389. Synthesis of (1r,2R,3S)—N-((6³S,3S,4S,Z)-3-ethoxy-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-((8S,9aS)-octahydropyrido[2,1-c][1,4]oxazin-8-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2,3-dimethylcyclopropane-1-carboxamide

Step 1.

To a mixture of(6³S,3S,4S,Z)-4-amino-3-ethoxy-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-((8S,9aS)-octahydropyrido[2,1-c][1,4]oxazin-8-yl)pyridin-3-yl)-10,10-dimethyl-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-5,7-dioneHCl salt (90 mg, 0.11 mmol) and(1R,2R,3S)-2,3-dimethylcyclopropane-1-carboxylic acid (19 mg, 0.17 mmol)in DMF (3 mL) at 0° C. was added DIPEA (429 mg, 3.3 mmol) and HATU (63mg, 0.17 mmol) in portions. The mixture was warmed to room temperatureand stirred for 2 h, then diluted with H₂O (10 mL) and the mixture wasextracted with EtOAc (2×10 mL). The combined organic layers were washedwith brine (3×10 mL), then concentrated under reduced pressure and theresidue was purified by preparative-HPLC to give(1r,2R,3S)—N-((6³S,3S,4S,Z)-3-ethoxy-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-((8S,9aS)-octahydropyrido[2,1-c][1,4]oxazin-8-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2,3-dimethylcyclopropane-1-carboxamide(13.8 mg, 13% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₅₀H₆₇N₇O₇S. 909.5; found 910.7; ¹H NMR (400 MHz, DMSO-d₆) δ 8.77 (s,1H), 8.50 (s, 1H), 7.93 (s, 1H), 7.83-7.51 (m, 4H), 5.96-5.77 (m, 1H),5.17 (d, J=11.5 Hz, 1H), 4.92 (s, 1H), 4.45-4.03 (m, 5H), 3.57 (ddd,J=34.6, 24.2, 13.9 Hz, 8H), 3.23 (d, J=8.3 Hz, 4H), 3.08 (d, J=10.4 Hz,2H), 2.84 (d, J=45.9 Hz, 3H), 2.67 (s, 2H), 2.23 (s, 2H), 2.17-2.01 (m,7H), 1.81 (s, 3H), 1.53 (s, 4H), 1.45-1.30 (m, 4H), 1.24 (s, 1H), 1.16(td, J=7.0, 1.9 Hz, 4H), 1.06 (dd, J=12.1, 5.1 Hz, 6H), 0.97-0.76 (m,8H), 0.38 (s, 3H).

Example A390. Synthesis of(1r,2R,3S)—N-((6³S,3S,4S,Z)-3-ethoxy-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-((8R,9aS)-octahydropyrido[2,1-c][1,4]oxazin-8-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2,3-dimethylcyclopropane-1-carboxamide

Step 1.

To a mixture of(6³S,3S,4S,Z)-4-amino-3-ethoxy-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-((8R,9aS)-octahydropyrido[2,1-c][1,4]oxazin-8-yl)pyridin-3-yl)-10,10-dimethyl-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-5,7-dione(100 mg, 0.12 mmol) and(1R,2R,3S)-2,3-dimethylcyclopropane-1-carboxylicacid (21 mg, 0.18 mmol)in DMF (3 mL) at −10° C. under an atmosphere of N₂ was added DIPEA (476mg, 3.7 mmol) and HATU (56 mg, 0.15 mmol, 1.2 equiv) in portions. Themixture was stirred at −10° C. for 1.5 h, then diluted with brine (5 mL)and extracted with EtOAc (3×10 mL). The combined organic layers werewashed with brine (3×10 mL), dried over anhydrous Na₂SO₄ and thefiltered. The filtrate was concentrated under reduced pressure and theresidue was purified by preparative-HPLC to give(1r,2R,3S)—N-((6³S,3S,4S,Z)-3-ethoxy-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-((8R,9aS)-octahydropyrido[2,1-c][1,4]oxazin-8-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2,3-dimethylcyclopropane-1-carboxamide(22 mg, 20% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₅₀H₆₇N₇O₇S. 909.5; found 910.7; ¹H NMR (400 MHz, DMSO-d₆) δ 8.65 (d,J=2.1 Hz, 1H), 8.49 (s, 1H), 7.92 (s, 1H), 7.74 (d, J=8.5 Hz, 1H),7.68-7.54 (m, 3H), 5.88 (d, J=9.8 Hz, 1H), 5.17 (d, J=12.1 Hz, 1H), 4.93(s, 1H), 4.36-3.99 (m, 5H), 3.80-3.43 (m, 7H), 3.24-3.05 (m, 4H),2.92-2.72 (m, 4H), 2.70-2.59 (m, 1H), 2.28-1.99 (m, 5H), 1.89-1.59 (m,6H), 1.52 (q, J=8.2, 6.3 Hz, 2H), 1.42-1.22 (m, 5H), 1.20-0.99 (m, 12H),0.86 (d, J=25.0 Hz, 7H), 0.39 (s, 3H).

Example A391. Synthesis of(1r,2R,3S)—N-((6³S,4S,Z)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(2,4)-oxazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2,3-dimethylcyclopropane-1-carboxamide

Step 1.

A mixture of benzyl4-(5-((6³S,4S,Z)-4-((tert-butoxycarbonyl)amino)-1¹-ethyl-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(2,4)-oxazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-1²-yl)-6-((S)-1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylate(150 mg, 0.16 mmol), paraformaldehyde (36 mg, 0.81 mmol) and Pd(OH)₂,30% weight on carbon (151 mg, 0.32 mmol) in MeOH (3 mL) was hydrogenatedat 30° C. for 2 h. The mixture was filtered through a pad of Celite andthe filtrate was concentrated under reduced pressure to give tert-butyl((6³S,4S,Z)-11-ethyl-1²-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(2,4)-oxazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(134 mg) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd for C₄₄H₆₀N₈O₇ 812.5;found 813.4.

Step 2.

To a mixture of tert-butyl((6³S,4S,Z)-11-ethyl-1²-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(2,4)-oxazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(134 mg, 0.17 mmol) in DCM (1.5 mL) at 0° C. was added TFA (1.50 mL) inportions. The mixture was warmed to room temperature and stirred for 2h, then concentrated under reduced pressure to give(6³S,4S,Z)-4-amino-1′-ethyl-1²-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(2,4)-oxazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-5,7-dione(318 mg) as an oil. LCMS (ESI): m/z [M+H]⁺ calc'd for C₃₉H₅₂N₈O₅ 712.4;found 713.4.

Step 3.

To a mixture of(6³S,4S,Z)-4-amino-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(2,4)-oxazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-5,7-dione (130 mg, 0.18 mmol),(1R,2R,3S)-2,3-dimethylcyclopropane-1-carboxylic acid (42 mg, 0.36 mmol)and DIPEA (236 mg, 1.8 mmol) in DMF (2 mL) at 0° C. was added COMU (117mg, 0.27 mmol) in portions. The mixture was warmed to room temperatureand stirred for 2h, then the residue was purified by preparative-HPLC togive(1r,2R,3S)—N-((6³S,4S,Z)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(2,4)-oxazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2,3-dimethylcyclopropane-1-carboxamide(28 mg, 19% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₄₅H₆₀N₈O₆ 808.5; found 809.8; ¹H NMR (300 MHz, DMSO-d₆) δ 8.66-8.45 (m,2H), 8.29 (d, J=8.0 Hz, 1H), 7.88 (s, 1H), 7.74 (d, J=9.4 Hz, 2H), 7.30(s, 1H), 5.81 (s, 1H), 5.00 (d, J=11.7 Hz, 1H), 4.53-4.06 (m, 5H),3.77-3.59 (m, 2H), 3.34 (s, 4H), 3.21 (s, 3H), 3.05-2.61 (m, 5H), 2.52(s, 4H), 2.28 (s, 3H), 2.05 (s, 1H), 1.83 (s, 1H), 1.56 (s, 2H),1.48-1.32 (m, 3H), 1.30-0.98 (m, 12H), 0.93 (s, 3H), 0.46 (s, 3H).

Example A396. Synthesis of(2S)—N-((6³S,3S,4S,Z)-3-ethoxy-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2-((R)-1-hydroxyethyl)azetidine-1-carboxamide

Step 1.

To a mixture of tert-butyl (S)-2-acetylazetidine-1-carboxylate (543 mg,1.96 mmol) in DCM (330 mL) at room temperature under an atmosphere of N₂was added BH₃-Me₂S (2.48 g, 32.6 mmol). To the above mixture was addedtert-butyl (2S)-2-acetylazetidine-1-carboxylate [Org. Lett. 2019, 22,9981-9984-see Supporting Information] (2.6 g, 13.0 mmol) dropwise over10 min. The resulting mixture was stirred for additional 2 h at roomtemperature. The reaction was monitored by LCMS. The reaction wasquenched with MeOH at 0° C. and was concentrated under reduced pressure.The residue was purified by silica gel column chromatography to givetert-butyl (S)-2-((R)-1-hydroxyethyl)azetidine-1-carboxylate (1.1 g, 42%yield) as an oil. LCMS (ESI): m/z [M-C₄H₈+H]⁺ calc'd for C₆H₁₁NO₃ 145.1;found 146.1.

Step 2.

To a mixture of tert-butyl(S)-2-((R)-1-hydroxyethyl)azetidine-1-carboxylate (300 mg, 1.49 mmol) inDCM (5 mL) at 0° C. was added 4M HCl in 1,4-dioxane (5 mL). The mixturewas stirred until completion, then concentrated under reduced pressureto give (R)-1-((S)-azetidin-2-yl)ethan-1-ol (310 mg) as a solid, whichwas used directly in the next step without further purification. LCMS(ESI): m/z [M+H]⁺ calc'd for C₅H₁₁NO 101.1; found 102.2.

Step 3.

To a mixture of(6³S,3S,4S,Z)-4-amino-3-ethoxy-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-5,7-dione(150 mg, 0.19 mmol), pyridine (1 mL), and DCM (2 mL) at 0° C. was added4-nitrophenyl chloroformate (65 mg, 0.39 mmol). The mixture was warmedto room temperature and stirred for 4 h, then concentrated under reducedpressure to give 4-nitrophenyl((6³S,3S,4S,Z)-3-ethoxy-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(crude) as an oil, which was used directly in the next step withoutfurther purification. LCMS (ESI): m/z [M+H]⁺ calc'd for C₄₈H₅₉N₉O₉S.937.4; found 938.3.

Step 4.

To a mixture of 4-nitrophenyl((6³S,3S,4S,Z)-3-ethoxy-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(195 mg, 0.21 mmol) and (R)-1-((S)-azetidin-2-yl)ethan-1-ol (285 mg, 2.8mmol) in MeCN (2 mL) at 0° C. was added DIPEA (403 mg, 3.1 mmol)dropwise. The mixture was warmed to room temperature and stirred for 1h. The residue was purified by preparative-HPLC to give(2S)—N-((6³S,3S,4S,Z)-3-ethoxy-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2-((R)-1-hydroxyethyl)azetidine-1-carboxamide(4.8 mg, 3%) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd for C₄₇H₆₅N₉O₇S.899.5; found 900.9; ¹H NMR (400 MHz, DMSO-d₆) δ 8.52-8.45 (m, 2H), 7.91(s, 1H), 7.76-7.72 (m, 1H), 7.58-7.53 (m, 1H), 7.22 (s, 1H), 6.73-6.68(m, 1H), 5.54-5.50 (m, 1H), 5.31-5.26 (m, 1H), 5.16-5.12 (m, 1H), 4.91(s, 1H), 4.30-4.24 (m, 3H), 4.16-4.04 (m, 3H), 3.84-3.67 (m, 2H),3.69-3.41 (m, 5H), 3.28-3.23 (m, 4H), 3.16 (s, 3H), 2.84-2.79 (m, 2H),2.48-2.44 (m, 4H), 2.23-2.21 (m, 4H), 2.08-2.05 (m, 1H), 1.93-1.89 (m,1H), 1.79-1.77 (m, 2H), 1.54-1.52 (m, 1H), 1.36-1.33 (m, 3H), 1.26-1.16(m, 4H), 1.13-1.10 (m, 3H), 1.00-0.73 (m, 6H), 0.43 (s, 2H).

Examples A424 and A441. Synthesis of(1R,2R,3S)—N-((2³S,6³S,4S)-1¹-ethyl-2³-hydroxy-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-2¹,2²,2³,2⁶,6¹,6²,6³,6⁴,6⁵,6⁶-decahydro-1¹H-8-oxa-1(5,3)-indola-6(1,3)-pyridazina-2(5,1)-pyridinacycloundecaphane-4-yl)-2,3-dimethylcyclopropane-1-carboxamideand(1R,2R,3S)—N-((2³R,6³S,4S)-1¹-ethyl-2³-hydroxy-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-2¹,2²,2³,2⁶,6¹,6²,6³,6⁴,6⁵,6⁶-decahydro-1¹H-8-oxa-1(5,3)-indola-6(1,3)-pyridazina-2(5,1)-pyridinacycloundecaphane-4-yl)-2,3-dimethylcyclopropane-1-carboxamide

Step 1.

To a mixture of 1-benzyl-3,5-dibromo-1,2,3,6-tetrahydropyridine (40.0 g,120.8 mmol) in MeCN (900 mL) and H₂O (600 mL) at 0° C. was added NH₄HCO₃(14.33 g, 181.2 mmol). The mixture was warmed to room temperature andstirred for 16 h, then concentrated under reduced pressure and theresidue was purified by reverse-phase silica gel column chromatographyto give 1-benzyl-5-bromo-1,2,3,6-tetrahydropyridin-3-ol (16.0 g, 49%yield) as an oil. LCMS (ESI): m/z [M+H]⁺ calc'd for C₁₂H₁₄BrNO 267.0;found 268.1.

Step 2.

To a mixture of 1-benzyl-5-bromo-1,2,3,6-tetrahydropyridin-3-ol (15.0 g,55.9 mmol) in DCM (150 mL) was added TBDPSCl (23.1 g, 83.9 mmol) andimidazole (7.62 g, 111.9 mmol) under an atmosphere of N₂ for 16 h. H2Owas added and the mixture and was extracted with EtOAc (3×300 mL). Thecombined organic layers were washed with brine (3×300 mL), dried overanhydrous Na₂SO₄ and filtered. The filtrate was concentrated underreduced pressure and the residue was purified by reverse-phase silicagel column chromatography to give1-benzyl-5-bromo-3-((tert-butyldiphenylsilyl)oxy)-1,2,3,6-tetrahydropyridine(15.0 g, 53% yield) as an oil. LCMS (ESI): m/z [M+H]⁺ calc'd forC₂₈H₃₂BrNOSi 505.1; found 506.2.

Step 3.

To a mixture of1-benzyl-5-bromo-3-((tert-butyldiphenylsilyl)oxy)-1,2,3,6-tetrahydropyridine(15.0 g, 29.6 mmol) in DCM (150 mL) at 0° C. was added 2-chloroethylchloroformate (16.93 g, 118.5 mmol). The mixture was warmed to 40° C.and was stirred for 4 h, then diluted with H₂O and the mixture wasextracted with EtOAc (2×300 mL). The combined organic layers were washedwith brine (2×300 mL), dried over anhydrous Na₂SO₄ and filtered. Thefiltrate was concentrated under reduced pressure and the residue wasdissolved in MeOH (150 mL). The mixture was heated to 70° C. and stirredfor 2 h, then the combined organic layers were washed with NaHCO₃ (2×300mL), dried over anhydrous Na₂SO₄ and filtered. The filtrate wasconcentrated under reduced pressure and the residue was purified byreverse-phase silica gel column chromatography to give5-bromo-3-((tert-butyldiphenylsilyl)oxy)-1,2,3,6-tetrahydropyridine (9.0g, 73% yield) as an oil. LCMS (ESI): m/z [M+H]⁺ calc'd for C₂₁H₂₆BrNOSi417.1; found 418.0 [for ⁸¹Br].

Step 4.

To a mixture of5-bromo-3-((tert-butyldiphenylsilyl)oxy)-1,2,3,6-tetrahydropyridine (6.0g, 14.4 mmol) in MeCN (60 mL) and H₂O (60 mL) at 0° C. was addedtert-butyl (S)-(2-oxooxetan-3-yl)carbamate (2.97 g, 15.9 mmol) andCs₂CO₃ (11.74 g, 36.0 mmol). The mixture was warmed to 40° C. andstirred for 16 h, then extracted with EtOAc (2×100 mL). The combinedorganic layers were washed with brine (3×200 mL), dried over anhydrousNa₂SO₄ and filtered. The filtrate was concentrated under reducedpressure and the residue was purified by reverse-phase silica gel columnchromatography to give(2S)-3-(5-bromo-3-((tert-butyldiphenylsilyl)oxy)-3,6-dihydropyridin-1(2H)-yl)-2-((tert-butoxycarbonyl)amino)propanoicacid (4.0 g, 46% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₂₉H₃₉BrN₂O₅Si, 602.2; found 603.1.

Step 5.

To a mixture of3-(1-ethyl-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)-2,2-dimethylpropyl(S)-hexahydropyridazine-3-carboxylate (1.0 g, 1.7 mmol) in DMF (10 mL)at 0° C. was added DIPEA (1.07 g, 8.3 mmol),(2S)-3-(5-bromo-3-((tert-butyldiphenylsilyl)oxy)-3,6-dihydropyridin-1(2H)-yl)-2-((tert-butoxycarbonyl)amino)propanoic acid (749 mg, 1.24mmol) and HATU (1.26 g, 3.3 mmol). The mixture was stirred at 0° C. for2 h, then diluted with H2O, and the mixture was extracted with EtOAc(3×100 mL). The combined organic layers were washed with brine (3×100mL), dried over anhydrous Na₂SO₄ and filtered. The filtrate wasconcentrated under reduced pressure and the residue was purified bypreparative-TLC to give3-(1-ethyl-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)-2,2-dimethylpropyl-(3S)-1-((2S)-3-(5-bromo-3-((tert-butyldiphenylsilyl)oxy)-3,6-dihydropyridin-1(2H)-yl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylate(1.2 g, 61% yield) as an oil. LCMS (ESI): m/z [M+H]⁺ calc'd forC₆₃H₈₆BBrN₆O₉Si, 1188.6; found 1189.4.

Step 6.

To a mixture of3-(1-ethyl-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)-2,2-dimethylpropyl-(3S)-1-((2S)-3-(5-bromo-3-((tert-butyldiphenylsilyl)oxy)-3,6-dihydropyridin-1(2H)-yl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylate(1.2 g, 1.0 mmol) in toluene (30 mL), 1,4-dioxane (10 mL) and H₂O (10mL) was added K₂CO₃ (418 mg, 3.0 mmol) and Pd(dppf)Cl₂ (74 mg, 0.1mmol). The mixture was heated to 65° C. and stirred for 2 h, thendiluted with H₂O, and the mixture was extracted with EtOAc (2×100 mL).The combined organic layers were washed with brine (2×100 mL), driedover anhydrous Na₂SO₄ and filtered. The filtrate was concentrated underreduced pressure and the residue was purified by preparative-TLC to givetert-butyl((6³S,4S)-2³-((tert-butyldiphenylsilyl)oxy)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-2¹,2²,2³,2⁶,6¹,6²,6³,6⁴,6⁵,6⁶-decahydro-1¹H-8-oxa-1(5,3)-indola-6(1,3)-pyridazina-2(5,1)-pyridinacycloundecaphane-4-yl)carbamate (420 mg, 42% yield) as an oil.LCMS (ESI): m/z [M+H]⁺ calc'd for C₅₇H₇₄N₆O₇Si, 982.5; found 984.1.

Step 7.

To a mixture of tert-butyl((6³S,4S)-2³-((tert-butyldiphenylsilyl)oxy)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-2¹,2²,2³,2⁶,6¹,6²,6³,6⁴,6⁵,6⁶-decahydro-1¹H-8-oxa-1(5,3)-indola-6(1,3)-pyridazina-2(5,1)-pyridinacycloundecaphane-4-yl)carbamate(420 mg, 0.43 mmol) in DCM (10 mL) at 0° C. was added ZnBr₂ (481 mg,2.14 mmol). The mixture was warmed to room temperature and stirred for16 h, then filtered, and the filter cake was washed with EtOAc (3×20mL). The filtrate was concentrated under reduced pressure to give(6³S,4S)-4-amino-2³-((tert-butyldiphenylsilyl)oxy)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-2¹,2²,2³,2⁶,6¹,6²,6³,6⁴,6⁵,6⁶-decahydro-1¹H-8-oxa-1(5,3)-indola-6(1,3)-pyridazina-2(5,1)-pyridinacycloundecaphane-5,7-dione (400 mg) which was used directly inthe next step without further purification. LCMS (ESI): m/z [M+H]⁺calc'd for C₅₂H₆₆N₆O₅Si, 882.5; found 883.6.

Step 8.

To a mixture of(6³S,4S)-4-amino-2³-((tert-butyldiphenylsilyl)oxy)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-2¹,2²,2³,2⁶,6¹,6²,6³,6⁴,6⁵,6⁶-decahydro-1¹H-8-oxa-1(5,3)-indola-6(1,3)-pyridazina-2(5,1)-pyridinacycloundecaphane-5,7-dione(380 mg, 0.43 mmol) in DMF (4 mL) at 0° C. was added DIPEA (556 mg, 4.3mmol), (1R,2R,3S)-2,3-dimethylcyclopropane-1-carboxylic acid (74 mg,0.65 mmol) and HATU (327 mg, 0.86 mmol). The mixture was stirred at 0°C. for 2 h, then diluted with H₂O, and the mixture was extracted withEtOAc (3×50 mL). The combined organic layers were washed with brine(3×50 mL), dried over anhydrous Na₂SO₄ and filtered. The filtrate wasconcentrated under reduced pressure and the residue was purified bypreparative-TLC to give(1R,2R,3S)—N-((6³S,4S)-2³-((tert-butyldiphenylsilyl)oxy)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-2¹,2²,2³,2⁶,6¹,6²,6³,6⁴,6⁵,6⁶-decahydro-1¹H-8-oxa-1(5,3)-indola-6(1,3)-pyridazina-2(5,1)-pyridinacycloundecaphane-4-yl)-2,3-dimethylcyclopropane-1-carboxamide(190 mg, 45% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₅₈H₇₄N₆O₆Si, 978.5; found 979.7.

Step 9.

To a mixture of(1R,2R,3S)—N-((6³S,4S)-2³-((tert-butyldiphenylsilyl)oxy)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-2¹,2²,2³,2⁶,6¹,6²,6³,6⁴,6⁵,6⁶-decahydro-1¹H-8-oxa-1(5,3)-indola-6(1,3)-pyridazina-2(5,1)-pyridinacycloundecaphane-4-yl)-2,3-dimethylcyclopropane-1-carboxamide(190 mg, 0.19 mmol) in THF (4 mL) at 0° C. was added TBAF (811 mg, 3.1mmol) and AcOH (1 mg, 0.02 mmol). The mixture was warmed to roomtemperature and stirred for 16 h, then diluted with H₂O, and the mixturewas extracted with EtOAc (3×50 mL). The combined organic layers werewashed with brine (3×50 mL), dried over anhydrous Na₂SO₄ and filtered.The filtrate was concentrated under reduced pressure and the residue waspurified by preparative-HPLC to give(1R,2R,3S)—N-((2³S,6³S,4S)-1¹-ethyl-2³-hydroxy-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-2¹,2²,2³,2⁶,6¹,6²,6³,6⁴,6⁵,6⁶-decahydro-1¹H-8-oxa-1(5,3)-indola-6(1,3)-pyridazina-2(5,1)-pyridinacycloundecaphane-4-yl)-2,3-dimethylcyclopropane-1-carboxamide(26 mg, 13% yield) and(1R,2R,3S)—N-((2³R,6³S,4S)-1¹-ethyl-2³-hydroxy-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-2¹,2²,2³,2⁶,6¹,6²,6³,6⁴,6⁵,6⁶-decahydro-1¹H-8-oxa-1(5,3)-indola-6(1,3)-pyridazina-2(5,1)-pyridinacycloundecaphane-4-yl)-2,3-dimethylcyclopropane-1-carboxamide(11 mg, 8% yield) both as solids. LCMS (ESI): m/z [M+H]⁺ calc'd forC₄₂H₅₆N₆O₆ 740.4; found 741.3; ¹H NMR (400 MHz, DMSO-d₆) δ 8.76 (dd,J=4.8, 1.7 Hz, 1H), 8.07 (d, J=8.4 Hz, 1H), 7.88 (dd, J=7.8, 1.8 Hz,1H), 7.61-7.55 (m, 1H), 7.55-7.47 (m, 2H), 7.42 (s, 1H), 6.24 (s, 1H),5.66 (d, J=12.2 Hz, 1H), 5.59 (t, J=9.1 Hz, 1H), 4.91 (d, J=6.7 Hz, 1H),4.40-4.26 (m, 2H), 4.13 (q, J=6.2 Hz, 1H), 4.08-3.96 (m, 2H), 3.88-3.78(m, 1H), 3.72 (t, J=11.5 Hz, 1H), 3.64 (q, J=11.0 Hz, 2H), 3.11 (d,J=14.2 Hz, 1H), 2.96 (dt, J=14.4, 7.1 Hz, 2H), 2.80 (d, J=23.8 Hz, 6H),2.15 (t, J=9.5 Hz, 1H), 2.06 (d, J=8.6 Hz, 1H), 1.94 (d, J=11.0 Hz, 1H),1.84-1.73 (m, 1H), 1.66-1.51 (m, 2H), 1.41 (d, J=6.2 Hz, 3H), 1.24 (s,1H), 1.19-1.05 (m, 7H), 1.05-0.96 (m, 6H), 0.88 (s, 3H), 0.48 (s, 3H)and LCMS (ESI): m/z [M+H]⁺ calc'd for C₄₂H₅₆N₆O₆ 740.4; found 741.4; ¹HNMR (400 MHz, DMSO-d₆) δ 8.76 (dd, J=4.7, 1.8 Hz, 1H), 8.12 (d, J=8.5Hz, 1H), 7.85 (dd, J=7.8, 1.7 Hz, 1H), 7.51 (q, J=4.5 Hz, 3H), 7.45 (s,1H), 6.19 (s, 1H), 5.70 (t, J=8.7 Hz, 1H), 5.49 (s, 1H), 4.72 (d, J=6.4Hz, 1H), 4.31 (d, J=12.7 Hz, 1H), 4.18 (d, J=6.2 Hz, 2H), 4.14-4.06 (m,1H), 3.99-3.68 (m, 3H), 3.61 (d, J=11.0 Hz, 1H), 3.53 (d, J=10.9 Hz,1H), 3.14 (d, J=15.8 Hz, 1H), 2.94 (s, 3H), 2.92-2.83 (m, 2H), 2.73 (d,J=14.1 Hz, 3H), 2.33 (q, J=1.8 Hz, 1H), 1.95 (d, J=10.5 Hz, 1H), 1.77(d, J=10.6 Hz, 1H), 1.64-1.48 (m, 2H), 1.40 (d, J=6.2 Hz, 3H), 1.24 (s,1H), 1.19-0.95 (m, 12H), 0.85 (s, 1H), 0.72 (s, 3H), 0.58 (s, 3H).

Example A437. Synthesis of(2S,6S)—N-((6³S,3S,4S,Z)-3-(2-(dimethylamino)ethoxy)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2,4,6-trimethylpiperazine-1-carboxamide

Step 1.

To a mixture of ethyl(2S,3S)-3-(4-bromothiazol-2-yl)-2-((tert-butoxycarbonyl)amino)-3-(2-oxoethoxy)propanoate(0.31 g, 6.9 mmol) and dimethylamine, 30% in MeOH (865 mg, 8.5 mmol) inMeOH (20 mL) at 0° C. was added NaBH₃CN (1.08 g, 17.2 mmol) over 2 min.The mixture was stirred at rt, then diluted with H₂O (30 mL) andextracted with EtOAc (3×20 mL). The combined organic layers were washedwith H₂O (4×10 mL), dried over anhydrous Na₂SO₄ and filtered. Thefiltrate was concentrated under reduced pressure and the residue waspurified by silica gel column chromatography to give ethyl(2S,3S)-3-(4-bromothiazol-2-yl)-2-((tert-butoxycarbonyl)amino)-3-(2-(dimethylamino)ethoxy)propanoate(1.2 g, 45% yield) as an oil. LCMS (ESI): m/z [M+H]⁺ calc'd forC₁₇H₂₈BrN₃O₅S. 467.1; found 468.2.

Step 2.

To a mixture of ethyl(2S,3S)-3-(4-bromothiazol-2-yl)-2-((tert-butoxycarbonyl)amino)-3-(2-(dimethylamino)ethoxy)propanoate(1.25 g, 2.68 mmol) in THF (9 mL) at 0° C. under an atmosphere of N₂ wasadded 1M LiOH (8.0 mL, 8.0 mmol) dropwise. The mixture was stirred at 0°C. for 2 h, then acidified to pH ˜6 with 1M HCl, then extracted withEtOAc (3×50 mL). The combined organic layers were washed with brine(3×10 mL), dried over anhydrous Na₂SO₄, filtered and the filtrate wasconcentrated under reduced pressure to give(2S,3S)-3-(4-bromothiazol-2-yl)-2-((tert-butoxycarbonyl)amino)-3-(2-(dimethylamino)ethoxy)propanoicacid (600 mg, 51% yield) as an oil. LCMS (ESI): m/z [M+H]⁺ calc'd forC₁₅H₂₄BrN₃O₅S. 439.1; found 440.3 [for ⁸¹Br].

Step 3.

A mixture of(2S,3S)-3-(4-bromothiazol-2-yl)-2-((tert-butoxycarbonyl)amino)-3-(2-(dimethylamino)ethoxy)propanoicacid (570 mg, 1.3 mmol),2-{[(2/W)-1-ethyl-2-{2-[(1S)-1-methoxyethyl]pyridin-3-yl}-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indol-3-yl]methyl}-2-methylpropyl(3S)-1,2-diazinane-3-carboxylate (1.18 g, 1.95 mmol) and DIPEA (5.04 g,39.0 mmol) in MECN (6 mL) at 0° C. under an atmosphere of N₂ was stirredfor 5 min, then HATU (593 mg, 1.56 mmol) was added in portions over 2min. The mixture was warmed to room temperature and stirred for 4 h,then diluted with H₂O (30 mL) and extracted with EtOAc (3×20 mL). Thecombined organic layers were washed with H₂O (3×10 mL), dried overanhydrous Na₂SO₄ and filtered. The filtrate was concentrated underreduced pressure and the residue was purified by preparative-TLC to give3-(1-ethyl-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)-2,2-dimethylpropyl-(S)-1-((2S,3S)-3-(4-bromothiazol-2-yl)-2-((tert-butoxycarbonyl)amino)-3-(2-(dimethylamino)ethoxy)propanoyl)hexahydropyridazine-3-carboxylate(580 mg, 44% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₄₉H₇₁BBrN₇O₉S. 1025.4; found 1026.5 [for ⁸¹Br].

Step 4.

To a mixture of3-(1-ethyl-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)-2,2-dimethylpropyl-(S)-1-((2S,3S)-3-(4-bromothiazol-2-yl)-2-((tert-butoxycarbonyl)amino)-3-(2-(dimethylamino)ethoxy)propanoyl)hexahydropyridazine-3-carboxylate(580 mg, 0.57 mmol) and K₃PO₄ (300 mg, 1.42 mmol) in toluene (6 mL),1,4-dioxane (2 mL) and H₂O (2 mL) under an atmosphere of Ar was addedPd(dtbpf)Cl₂. The mixture was heated to 60° C. and stirred for 4 h, thendiluted with H₂O (30 mL), and the mixture was extracted with EtOAc (3×30mL). The combined organic layers were washed with brine (3×10 mL), driedover anhydrous Na₂SO₄ and filtered. The filtrate was concentrated underreduced pressure and the residue was purified by preparative-TLC to give(2S,6S)—N-((6³S,3S,4S,Z)-3-(2-(dimethylamino)ethoxy)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2,4,6-trimethylpiperazine-1-carboxamide(230 mg, 50% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₄₃H₅₉N₇O₇S. 817.4; found 818.4.

Step 5.

To a mixture of tert-butyl((6³S,3S,4S,Z)-3-(2-(dimethylamino)ethoxy)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(230 mg, 0.28 mmol) in DCM (3 mL) at 0° C. under an atmosphere of N₂ wasadded HCl in 1,4-dioxane (1 mL). The mixture was warmed to roomtemperature and stirred for 2 h, then concentrated under reducedpressure, toluene (30 mL) was added to the residue, and the mixture wasconcentrated under reduced pressure to give(6³S,3S,4S,Z)-4-amino-3-(2-(dimethylamino)ethoxy)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-5,7-dione(200 mg) as a solid, which was used directly in the next step withoutfurther purification. LCMS (ESI): m/z [M+H]⁺ calc'd for C₃₈H₅₁N₇O₅S.717.4; found 718.7.

Step 6.

To a mixture of(6³S,3S,4S,Z)-4-amino-3-(2-(dimethylamino)ethoxy)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-5,7-dione(200 mg, 0.28 mmol) in THF (2 mL) at 0° C. was added NEt₃ (85 mg, 0.84mmol) dropwise over 1 min. The mixture was stirred at 0° C. for 5 min,then 4-nitrophenyl chloroformate (56 mg, 0.28 mmol) was added over 1min. The mixture was warmed to room temperature and stirred for 4 h andthe mixture was concentrated under reduced pressure to give4-nitrophenyl((6³S,3S,4S,Z)-3-(2-(dimethylamino)ethoxy)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate (250 mg) as a solid, that wasused directly in the next step without further purification.

Step 7.

To a mixture of 4-nitrophenyl((6³S,3S,4S,Z)-3-(2-(dimethylamino)ethoxy)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(200 mg, 0.23 mmol) in MeCN (2 mL) at 0° C. was added DIPEA (88 mg, 0.68mmol) dropwise. The mixture was stirred at 0° C. for 2 min, thentert-butyl (3S,5S)-3,5-dimethylpiperazine-1-carboxylate (97 mg, 0.45mmol) was added dropwise. The mixture was warmed to room temperature andstirred overnight, then diluted with H₂O (3 mL), and the mixture wasextracted with EtOAc (3×10 mL). The combined organic layers were washedwith brine (3×10 mL), dried over anhydrous Na₂SO₄ and filtered. Thefiltrate was concentrated under reduced pressure and the residue waspurified by preparative-TLC to give tert-butyl(3S,5S)-4-(((6³S,3S,4S,Z)-3-(2-(dimethylamino)ethoxy)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamoyl)-3,5-dimethylpiperazine-1-carboxylate(50 mg, 23% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₅₀H₇₁N₉O₈S. 957.5; found 958.9.

Step 8.

To a mixture of tert-butyl(3S,5S)-4-(((6³S,3S,4S,Z)-3-(2-(dimethylamino)ethoxy)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamoyl)-3,5-dimethylpiperazine-1-carboxylate(50 mg, 0.05 mmol) in DCM (3 mL) at 0° C. under an atmosphere of N₂ wasadded HCl in 1,4-dioxane (1 mL) dropwise. The mixture was warmed to roomtemperature and stirred for 2 h, then concentrated under reducedpressure, toluene (30 mL) was added to the residue and the mixture wasconcentrated under reduced pressure to give(2S,6S)—N-((6³S,3S,4S,Z)-3-(2-(dimethylamino)ethoxy)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2,6-dimethylpiperazine-1-carboxamide(50 mg) as a solid, that was used directly in the next step withoutfurther purification.

Step 9.

To a mixture of(2S,6S)—N-((6³S,3S,4S,Z)-3-(2-(dimethylamino)ethoxy)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2,6-dimethylpiperazine-1-carboxamide(40 mg, 0.05 mmol) and MeOH at 0° C. was added paraformaldehyde (11 mg,0.24 mmol) in portions, followed by NaBH₃CN (4.4 mg, 0.07 mmol) inportions. The mixture was warmed to room temperature and stirredovernight, then diluted with H₂O (5 mL) and extracted with EtOAc (3×10mL). The combined organic layers were washed with H₂O (3×10 mL), driedover anhydrous Na₂SO₄ and filtered. The filtrate was concentrated underreduced pressure and the residue was purified by preparative-HPLC togive(2S,6S)—N-((6³S,3S,4S,Z)-3-(2-(dimethylamino)ethoxy)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2,4,6-trimethylpiperazine-1-carboxamide(3.3 mg, 8% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₄₆H₆₅N₉O₆S. 871.5; found 872.3; ¹H NMR (400 MHz, DMSO-d₆) δ 8.74-8.64(dd, J=4.7, 1.6 Hz, 1H), 8.49-8.36 (s, 1H), 7.91-7.81 (s, 1H), 7.79-7.71(d, J=7.7 Hz, 1H), 7.71-7.63 (d, J=8.6 Hz, 1H), 7.56-7.47 (s, 1H),7.47-7.36 (dd, J=7.7, 4.7 Hz, 1H), 6.32-6.21 (d, J=9.9 Hz, 1H),5.55-5.46 (s, 1H), 5.19-5.11 (s, 1H), 5.04-4.97 (s, 1H), 4.45-3.94 (m,6H), 3.67-3.36 (m, 11H), 3.23 (s, 3H), 3.11 (s, 3H), 2.78-2.75 (m, 2H),2.40-2.34 (m, 3H), 2.15 (s, 6H), 2.11-2.04 (m, 6H), 1.75 (s, 2H),1.59-1.41 (m, 1H), 1.30 (d, J=6.0 Hz, 3H), 1.13 (s, 1H), 1.12 (d, J=6.1Hz, 6H), 0.80 (d, J=28.4 Hz, 6H), 0.57-0.12 (s, 3H).

Example A438. Synthesis of(1r,2R,3S)—N-((6³S,3S,4S,Z)-3-ethoxy-1¹-(2-isopropoxyethyl)-1²-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2,3-dimethylcyclopropane-1-carboxamide

Step 1.

To a mixture of5-bromo-3-{3-[(tert-butyldiphenylsilyl)oxy]-2,2-dimethylpropyl}-2-iodo-1H-indole(22.0 g, 34.0 mmol) and benzyl4-[6-(methoxymethyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-3-yl]piperazine-1-carboxylate(19.1 g, 40.8 mmol) in 1,4-dioxane (400 mL) and H₂O (80 mL) under anatmosphere of N₂ was added Pd(dppf)Cl₂ (2.49 g, 3.4 mmol) and K₂CO₃(11.76 g, 85.1 mmol) in portions. The mixture was heated to 70° C. andstirred for 16 h, then H₂O added, and the mixture was extracted withEtOAc (3×200 mL). The combined organic layers were washed with brine(3×100 mL), dried over anhydrous Na₂SO₄ and filtered. The filtrate wasconcentrated under reduced pressure and the residue was purified bysilica gel column chromatography to give benzyl(S)-4-(5-(5-bromo-3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-1H-indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylate(20 g, 67% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₄₉H₅₇BrN₄O₄Si, 874.3; found 875.5.

Step 2.

To a mixture of benzyl(S)-4-(5-(5-bromo-3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-1H-indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylate(23.0 g, 26.3 mmol) and 2-isopropoxyethyl 4-methylbenzenesulfonate (13.6g, 52.6 mmol) in DMF (300 mL) under an atmosphere of N₂ was added Cs₂CO₃(25.72 g, 79.0 mmol) in portions. The mixture was heated to 60° C. andstirred for 2, then diluted with brine (100 mL) and extracted with EtOAc(3×200 mL). The combined organic layers were washed with brine (3×200mL), dried over anhydrous Na₂SO₄ and filtered. The filtrate wasconcentrated under reduced pressure and the residue was purified bysilica gel column chromatography to give benzyl(S)-4-(5-(5-bromo-3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-1-(2-isopropoxyethyl)-1H-indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylate(19.2 g, 76% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₅₄H₆₇BrN₄O₅Si, 960.4; found 961.4 [for ⁸¹Br].

Step 3.

To a mixture of benzyl(S)-4-(5-(5-bromo-3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-1-(2-isopropoxyethyl)-1H-indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylate(3.0 g, 3.1 mmol) in THF (30 mL) at 0° C. under an atmosphere of N₂ wasadded TBAF, 1M in THF (15.6 mL, 15.6 mmol) in portions. The mixture washeated to 45° C. and stirred overnight, then diluted with brine (30 mL)and extracted with EtOAc (3×100 mL). The combined organic layers werewashed with brine (3×50 mL), dried over anhydrous Na₂SO₄ and filtered.The filtrate was concentrated under reduced pressure and the residue waspurified by silica gel column chromatography to give benzyl(S)-4-(5-(5-bromo-3-(3-hydroxy-2,2-dimethylpropyl)-1-(2-isopropoxyethyl)-1H-indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylate(1.19 g, 53% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₃₈H₄₉BrN₄O₅ 720.3; found 721.3.

Step 4.

To a mixture of benzyl(S)-4-(5-(5-bromo-3-(3-hydroxy-2,2-dimethylpropyl)-1-(2-isopropoxyethyl)-1H-indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylate(1.09 g, 1.51 mmol) and bis(pinacolato)diboron (0.58 g, 2.27 mmol) intoluene (11 mL) under an atmosphere of N₂ was added Pd(dppf)Cl₂ (0.11 g,0.15 mmol) and KOAc (0.37 g, 3.78 mmol) in portions. The mixture washeated to 80° C. and stirred for 2 h, then diluted with brine (10 mL)and extracted with EtOAc (3×20 mL). The combined organic layers werewashed with brine (3×20 mL), dried over anhydrous Na₂SO₄ and filtered.The filtrate was concentrated under reduced pressure and the residue waspurified by silica gel column chromatography to give benzyl(S)-4-(5-(3-(3-hydroxy-2,2-dimethylpropyl)-1-(2-isopropoxyethyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylate(1.0 g, 86% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₄₄H₆₁BN₄O₇ 768.5; found 769.7.

Step 5.

To a mixture of benzyl(S)-4-(5-(3-(3-hydroxy-2,2-dimethylpropyl)-1-(2-isopropoxyethyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylate(1.0 g, 1.3 mmol) and(3S)-1,2-bis(tert-butoxycarbonyl)-1,2-diazinane-3-carboxylic acid (0.64g, 2.0 mmol) in DCM (10 mL) at 0° C. under an atmosphere of N₂ was addedDMAP (0.24 g, 2.0 mmol) and DCC (0.40 g, 2.0 mmol) in portions. Themixture was warmed to room temperature and stirred overnight, thendiluted with brine (20 mL) and extracted with DCM (3×50 mL). Thecombined organic layers were washed with brine (3×20 mL), dried overanhydrous Na₂SO₄ and filtered. The filtrate was concentrated underreduced pressure and the residue was purified by silica gel columnchromatography to give3-(3-(2-(5-(4-((benzyloxy)carbonyl)piperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2-isopropoxyethyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)-2,2-dimethylpropyl)1,2-di-tert-butyl-(S)-tetrahydropyridazine-1,2,3-tricarboxylate(1.08 g, 77% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₅₉H₈₅BN₆O₁₂ 1080.6; found 1081.7.

Step 6.

To a mixture of3-(3-(2-(5-(4-((benzyloxy)carbonyl)piperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2-isopropoxyethyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)-2,2-dimethylpropyl)1,2-di-tert-butyl-(S)-tetrahydropyridazine-1,2,3-tricarboxylate(1.0 g, 0.9 mmol) in DCM (3 mL) at 0° C. under an atmosphere of N₂ wasadded HCl in 1,4-dioxane (9 mL) in portions. The mixture was warmed toroom temperature and stirred for 6 h, then diluted with toluene (30 mL)and concentrated under reduced pressure to give3-(2-(5-(4-((benzyloxy)carbonyl)piperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2-isopropoxyethyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)-2,2-dimethylpropyl-(S)-hexahydropyridazine-3-carboxylatebis hydrochloride (1.0 g) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₄₉H₆₉BN₆O₈ 880.5; found 881.5.

Step 7.

To a mixture of3-(2-(5-(4-((benzyloxy)carbonyl)piperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2-isopropoxyethyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)-2,2-dimethylpropyl-(S)-hexahydropyridazine-3-carboxylatebis hydrochloride (1.0 g, 1.1 mmol) and(2S,3S)-3-(4-bromo-1,3-thiazol-2-yl)-2-[(tert-butoxycarbonyl)amino]-3-ethoxypropanoicacid (0.54 g, 1.36 mmol) in DMF (10 mL) at 0° C. under an atmosphere ofN₂ was added DIPEA (4.40 g, 34.1 mmol) and (Z)-(ethylcyano({[(dimethyliminiumyl)(morpholin-4-yl)methoxy]imino})formate);hexafluorophosphate (0.53 g, 1.3 mmol) in portions. The mixture wasstirred at 0° C. for 1 h, then diluted with brine (30 mL) and extractedwith EtOAc (3×50 mL). The combined organic layers were washed with brine(3×30 mL), dried over anhydrous Na₂SO₄ and filtered. The filtrate wasconcentrated under reduced pressure and the residue was purified bypreparative-TLC to give3-(2-(5-(4-((benzyloxy)carbonyl)piperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2-isopropoxyethyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)-2,2-dimethylpropyl-(S)-1-((2S,3S)-3-(4-bromothiazol-2-yl)-2-((tert-butoxycarbonyl)amino)-3-ethoxypropanoyl)hexahydropyridazine-3-carboxylate(1.0 g, 70% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₆₂H₈₆BBrN₈O₁₂S. 1258.5; found 1259.5.

Step 8.

To a mixture of3-(2-(5-(4-((benzyloxy)carbonyl)piperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2-isopropoxyethyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)-2,2-dimethylpropyl-(S)-1-((2S,3S)-3-(4-bromothiazol-2-yl)-2-((tert-butoxycarbonyl)amino)-3-ethoxypropanoyl)hexahydropyridazine-3-carboxylate(1.0 g, 0.8 mmol) in toluene (30 mL), 1,4-dioxane (10 mL) and H₂O (10mL) under an atmosphere of N₂ was added Pd(dtbpf)Cl₂ (0.16 g, 0.24 mmol)and K₃PO₄ (0.42 g, 2.0 mmol) in portions. The mixture was heated to 65°C. and stirred for 2 h, then diluted with H₂O and extracted with EtOAc(3×50 mL). The combined organic layers were washed with brine (3×50 mL),dried over anhydrous Na₂SO₄ and filtered. The filtrate was concentratedunder reduced pressure and the residue was purified by preparative-TLCto give benzyl4-(5-((6³S,3S,4S,Z)-4-((tert-butoxycarbonyl)amino)-3-ethoxy-1¹-(2-isopropoxyethyl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-1²-yl)-6-((S)-1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylate(370 mg, 44% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₅₆H₇₄N₈O₁₀S. 1050.5; found 1051.9.

Step 9.

To a mixture of benzyl4-(5-((6³S,3S,4S,Z)-4-((tert-butoxycarbonyl)amino)-3-ethoxy-1¹-(2-isopropoxyethyl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-1²-yl)-6-((S)-1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylate(340 mg, 0.32 mmol) in MeOH (5 mL) was added paraformaldehyde (49 mg,1.6 mmol) and Pd/C (34 mg, 0.32 mmol) in portions. The mixture wasstirred under an atmosphere of H₂ overnight, then filtered through a padof Celite and the filter cake was washed with MeOH (3×20 mL). Thefiltrate was concentrated under reduced pressure and the residue waspurified by preparative-TLC to give tert-butyl((6³S,3S,4S,Z)-3-ethoxy-1¹-(2-isopropoxyethyl)-1²-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(240 mg, 80% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₄₉H₇₀N₈O₈S. 930.5; found 931.5.

Step 10.

To a mixture of tert-butyl((6³S,3S,4S,Z)-3-ethoxy-1¹-(2-isopropoxyethyl)-1²-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(240 mg, 0.26 mmol) in DCM (2 mL) at 0° C. under an atmosphere of N₂ wasadded HCl in 1,4-dioxane (6 mL) in portions. The mixture was stirred at0° C. for 1 h, then diluted with toluene (20 mL) and concentrated underreduced pressure to give(6³S,3S,4S,Z)-4-amino-3-ethoxy-1¹-(2-isopropoxyethyl)-1²-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-5,7-dioneHCl salt (240 mg) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₄₄H₆₂N₈O₆S. 830.5; found 831.4.

Step 11.

To a mixture of(6³S,3S,4S,Z)-4-amino-3-ethoxy-1¹-(2-isopropoxyethyl)-1²-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-5,7-dioneHCl salt (100 mg, 0.12 mmol) and(1R,2R,3S)-2,3-dimethylcyclopropane-1-carboxylic acid (21 mg, 0.18 mmol)in DMF (5 mL) at 0° C. under an atmosphere of N₂ was added DIPEA (467mg, 3.6 mmol) and (Z)-(ethylcyano({[(dimethyliminiumyl)(morpholin-4-yl)methoxy]imino})formate);hexafluorophophate (62 mg, 0.14 mmol) in portions. The mixture wasstirred at 0° C. for 1 h, then diluted with brine (5 mL) and extractedwith EtOAc (3×5 mL). The combined organic layers were washed with brine(3×15 mL), dried over anhydrous Na₂SO₄ and filtered. The filtrate wasconcentrated under reduced pressure and the residue was purified by(1r,2R,3S)—N-((6³S,3S,4S,Z)-3-ethoxy-1¹-(2-isopropoxyethyl)-1²-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2,3-dimethylcyclopropane-1-carboxamide(38 mg, 34% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₅₀H₇₀N₈O₇S. 926.5; found 927.5; ¹H NMR (400 MHz, DMSO-d₆) δ 8.52-8.42(m, 2H), 7.91 (s, 1H), 7.72 (dd, J=8.5, 1.6 Hz, 1H), 7.61 (dd, J=18.7,9.2 Hz, 2H), 7.21 (d, J=2.9 Hz, 1H), 5.88 (d, J=9.8 Hz, 1H), 5.19 (d,J=12.2 Hz, 1H), 4.92 (s, 1H), 4.41-4.02 (m, 5H), 3.73-3.47 (m, 4H), 3.25(q, J=4.1, 2.7 Hz, 8H), 3.13 (s, 3H), 2.78 (t, J=11.3 Hz, 2H), 2.49-2.39(m, 6H), 2.22 (s, 4H), 2.11-1.96 (m, 1H), 1.78 (d, J=29.7 Hz, 3H), 1.51(s, 3H), 1.32 (d, J=6.1 Hz, 4H), 1.24 (d, J=2.8 Hz, 1H), 1.20-0.97 (m,12H), 0.84 (dd, J=23.4, 6.0 Hz, 10H), 0.44 (s, 3H).

Example A449. Synthesis of(1r,2R,3S)—N-((6³S,3S,4S,Z)-3-ethoxy-1′-ethyl-1²-(2-((S)-1-methoxyethyl)-5-(1-(oxetan-3-yl)piperidin-4-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2,3-dimethylcyclopropane-1-carboxamide

Step 1.

A mixture of(2S,3S)-3-(4-bromothiazol-2-yl)-2-((tert-butoxycarbonyl)amino)-3-ethoxypropanoicacid (1.11 g, 7.7 mmol) and DIPEA (4.95 g, 38.3 mmol) in DMF (20 mL) wasstirred at room temperature for 5 min, then(2S,3S)-3-(4-bromo-1,3-thiazol-2-yl)-2-[(tert-butoxycarbonyl)amino]-3-ethoxypropanoicacid (1.52 g, 3.8 mmol) and HATU (2.91 g, 7.7 mmol) were added. When thereaction was complete by LCMS, it was cooled to 0° C., diluted with H₂Oand extracted with EtOAc (50 mL). The organic layer was washed withbrine (3×100 mL), dried over anhydrous Na₂SO₄ and filtered. The filtratewas concentrated under reduced pressure and the residue was purified bysilica gel column chromatography to give methyl(S)-1-((2S,3S)-3-(4-bromothiazol-2-yl)-2-((tert-butoxycarbonyl)amino)-3-ethoxypropanoyl)hexahydropyridazine-3-carboxylate (1.56 g, 78% yield). LCMS (ESI): m/z[M+H]⁺ calc'd for C₁₉H₂₉BrN₄O₆S. 522.1; found 523.1 [for ⁸¹Br],

Step 2.

A mixture of methyl(3S)-1-[(2S,3S)-3-(4-bromo-1,3-thiazol-2-yl)-2-[(tert-butoxycarbonyl)amino]-3-ethoxypropanoyl]-1,2-diazinane-3-carboxylate(1.56 g, 3.0 mmol), LiOH.H₂O (0.63 g, 15.0 mmol), THF (5 mL) and H₂O (5mL) at room temperature was stirred until complete by LCMS. The mixturewas acidified to pH ˜7 with 1N HCl was extracted with EtOAc (3×100 mL).The combined organic layers were dried over anhydrous Na₂SO₄, filtered,and the filtrate was concentrated under reduced pressure to give(3S)-1-[(2S,3S)-3-(4-bromo-1,3-thiazol-2-yl)-2-[(tert-butoxycarbonyl)amino]-3-ethoxypropanoyl]-1,2-diazinane-3-carboxylicacid (1.3 g), which was used directly in the next step without furtherpurification. LCMS (ESI): m/z [M+H]⁺ calc'd for C₁₉H₂₉BrN₄O₆S. 506.1;found 507.1.

Step 3.

A mixture of(3S)-1-[(2S,3S)-3-(4-bromo-1,3-thiazol-2-yl)-2-[(tert-butoxycarbonyl)amino]-3-ethoxypropanoyl]-1,2-diazinane-3-carboxylicacid (600 mg, 1.18 mmol), benzyl(5M)-5-[1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indol-2-yl]-6-[(1S)-1-methoxyethyl]-3′,6′-dihydro-2′H-[3,4′-bipyridine]-1′-carboxylate(754 mg, 1.1 mmol), DMAP (29 mg, 0.24 mmol), DCC (488 mg, 2.37 mmol) andDCM (15 mL) at room temperature until deemed complete by LCMS. Themixture was diluted with DCM (100 mL) and washed with H₂O (100 mL), thenthe aqueous layer was extracted with DCM (3×100 mL). The combinedorganic layers were dried over anhydrous Na₂SO₄, filtered, the filtratewas concentrated under reduced pressure and the residue was purified bysilica gel column chromatography to give benzyl(5M)-5-(3-{3-[(3S)-1-[(2S,3S)-3-(4-bromo-1,3-thiazol-2-yl)-2-[(tert-butoxycarbonyl)amino]-3-ethoxypropanoyl]-1,2-diazinane-3-carbonyloxy]-2,2-dimethylpropyl}-1-ethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indol-2-yl)-6-[(1S)-1-methoxyethyl]-3′,6′-dihydro-2′H-[3,4′-bipyridine]-1′-carboxylate(735 mg, 52% yield) as an oil. LCMS (ESI): m/z [M+H]⁺ calc'd forC₆₀H₇₉BBrN₇O₁₁S. 1195.5; found 1196.4.

Step 4.

A mixture of benzyl(5M)-5-(3-{3-[(3S)-1-[(2S,3S)-3-(4-bromo-1,3-thiazol-2-yl)-2-[(tert-butoxycarbonyl)amino]-3-ethoxypropanoyl]-1,2-diazinane-3-carbonyloxy]-2,2-dimethylpropyl}-1-ethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indol-2-yl)-6-[(1S)-1-methoxyethyl]-3′,6′-dihydro-2′H-[3,4′-bipyridine]-1′-carboxylate(725 mg, 0.61 mmol), K₃PO₄ (322 mg, 1.52 mmol), Pd(DtBPF)Cl₂ (79 mg,0.12 mmol), toluene (9 mL), 1,4-dioxane (3 mL) and H₂O (3 mL) under anatmosphere of N₂ was heated to 70° C. and stirred for 1 h at 70° C. Themixture was diluted with H₂O (50 mL) and extracted with EtOAc (3×100mL). The combined organic layers were dried over anhydrous Na₂SO₄,filtered, the filtrate was concentrated under reduced pressure and theresidue was purified by silica gel column chromatography to give benzyl5-((6³S,3S,4S,Z)-4-((tert-butoxycarbonyl)amino)-3-ethoxy-1′-ethyl-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-1²-yl)-6-((S)-1-methoxyethyl)-3′,6′-dihydro-[3,4′-bipyridine]-1′(2′H)-carboxylate(233 mg, 39% yield) as a an oil. LCMS (ESI): m/z [M+H]⁺ calc'd forC₅₄H₆₇N₇O₉S. 989.5; found 990.6.

Step 5.

A mixture of benzyl5-((6³S,3S,4S,Z)-4-((tert-butoxycarbonyl)amino)-3-ethoxy-1′-ethyl-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-1²-yl)-6-((S)-1-methoxyethyl)-3′,6′-dihydro-[3,4′-bipyridine]-1′(2′H)-carboxylate(223 mg, 0.23 mmol) and Pd(OH)₂/C (200 mg, 1.4 mmol) in MeOH (2 mL) wasstirred under an atmosphere of H₂ until deemed complete by LCMS. Themixture was filtered, and the filter cake was washed with MeOH (3×20mL). The filtrate was concentrated under reduced pressure to givetert-butyl((6³S,3S,4S,Z)-3-ethoxy-1′-ethyl-1²-(2-((S)-1-methoxyethyl)-5-(piperidin-4-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(192 mg) as a solid, that was used in the next step without furtherpurification. LCMS (ESI): m/z [M+H]⁺ calc'd for C₄₆H₆₃N₇O₇S. 875.5;found 858.4.

Step 6.

A mixture of tert-butyl((6³S,3S,4S,Z)-3-ethoxy-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-(piperidin-4-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(80 mg, 0.09 mmol), 3-oxetanone (134 mg, 1.86 mmol), AcOH (56 mg, 0.93mmol) and NaBH₃CN (59 mg, 0.93 mmol) in ^(i)PrOH (2 mL) at roomtemperature was stirred until deemed complete by LCMS. The mixture wascooled to 0° C. quenched with saturated NaHCO₃ and extracted with EtOAc(3×30 mL). The combined organic layers were dried over anhydrous Na₂SO₄,filtered, the filtrate was concentrated under reduced pressure and theresidue was purified by silica gel column chromatography to givetert-butyl((6³S,3S,4S,Z)-3-ethoxy-1′-ethyl-1²-(2-((S)-1-methoxyethyl)-5-(1-(oxetan-3-yl)piperidin-4-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(30 mg, 35% yield) as a an oil. LCMS (ESI): m/z [M+H]⁺ calc'd forC₄₉H₆₇N₇O₈S. 913.5; found 914.4.

Step 7.

A mixture of tert-butyl((6³S,3S,4S,Z)-3-ethoxy-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-(1-(oxetan-3-yl)piperidin-4-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(40 mg, 0.05 mmol), ZnBr₂(55 mg, 0.24 mmol) and DCM (1 mL) was stirredat room temperature until deemed complete by LCMS. The mixture wasfiltered, the filter cake was washed with DCM (3×10 mL), and thefiltrate was concentrated under reduced pressure to give(6³S,3S,4S,Z)-4-amino-3-ethoxy-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-(1-(oxetan-3-yl)piperidin-4-yl)pyridin-3-yl)-10,10-dimethyl-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-5,7-dioneas(crude)an oil, which was used directly in the next step without furtherpurification. LCMS (ESI): m/z [M+H]⁺ calc'd for C₄₄H₅₉N₇O₆S. 813.4;found 814.8.

Step 8.

To a mixture of(6³S,3S,4S,Z)-4-amino-3-ethoxy-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-(1-(oxetan-3-yl)piperidin-4-yl)pyridin-3-yl)-10,10-dimethyl-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-5,7-dioneas(40 mg, 0.05 mmol) in DMF (2 mL) at room temperature was added DIPEA (64mg, 0.49 mmol), (1R,2R,3S)-2,3-dimethylcyclopropane-1-carboxylic acid (6mg, 0.05 mmol) and HATU (37 mg, 0.1 mmol) in portions. The mixture wasstirred until deemed complete by LCMS. The mixture was diluted with H₂O(10 mL), extracted with EtOAc (50 mL), the organic layer was washed withbrine (3×50 mL), dried over anhydrous Na₂SO₄ and filtered. The filtratewas concentrated under reduced pressure and the residue was purified bypreparative-HPLC to give(1r,2R,3S)—N-((6³S,3S,4S,Z)-3-ethoxy-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-(1-(oxetan-3-yl)piperidin-4-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2,3-dimethylcyclopropane-1-carboxamide(3.5 mg, 8% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₅₀H₆₇N₇O₇S. 909.5; found 910.8; ¹H NMR (400 MHz, CD₃OD) δ 8.66-8.52 (m,2H), 7.82-7.72 (m, 2H), 7.67 (s, 1H), 7.53-7.44 (m, 1H), 5.99 (s, 1H),4.78-4.68 (m, 4H), 4.71-4.54 (m, 10H), 4.48-4.39 (m, 1H), 4.36-4.02 (m,4H), 3.75-3.68 (m, 1H), 3.68-3.64 (m, 3H), 3.02-2.97 (m, 3H), 2.91-2.80(m, 2H), 2.65-2.63 (m, 1H), 2.24-2.12 (m, 3H), 2.03-2.00 (m, 4H),1.93-1.72 (m, 3H), 1.69-1.55 (m, 1H), 1.47-1.23 (m, 6H), 1.16-1.13 (m,5H), 1.06-0.85 (m, 5H), 0.50 (s, 3H).

Example A451. Synthesis of(1S,2S)—N-((6³S,3S,4S,Z)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-3-((1-methylpiperidin-4-yl)methoxy)-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2-methylcyclopropane-1-carboxamide

Step 1.

To a mixture of methyl 1,2,3,6-tetrahydropyridine-4-carboxylate (10.0 g,70.8 mmol) and NaHCO₃ (29.75 g, 354.2 mmol) in THF (50 mL) and H₂O (50mL) was added CbzOSu (26.48 g, 106.3 mmol) in portions. The mixture wasstirred at room temperature for 2 h, then washed with H₂O (3×100 mL) andthe combined aqueous layers were extracted with EtOAc (3×100 mL). Thecombined organic layers were concentrated under reduced pressure and theresidue was purified by silica gel column chromatography to give1-benzyl 4-methyl 3,6-dihydropyridine-1,4(2H)-dicarboxylate as an oil.LCMS (ESI): m/z [M+H]⁺ calc'd for C₁₅H₁₇NO₄ 275.1; found 276.1.

Step 2.

To a mixture of 1-benzyl 4-methyl3,6-dihydropyridine-1,4(2H)-dicarboxylate (8.0 g, 29.1 mmol) in THF at−20° C. under an atmosphere of N₂ was added DIBAL-H (80 mL, 80 mmol)dropwise. The mixture was stirred at −20° C. for 2 h, then warmed toroom temperature and quenched with saturated NH₄Cl. The aqueous layerwas extracted with EtOAc (2×100 mL) and the combined organic layers wereconcentrated under reduced pressure. The residue was purified byreverse-phase silica gel column chromatography to give benzyl4-(hydroxymethyl)-3,6-dihydropyridine-1(2H)-carboxylate (2.3 g, 32%yield) as an oil. LCMS (ESI): m/z [M+H]⁺ calc'd for C₁₄H₁₇NO₃ 247.1;found 248.2.

Step 3.

To a mixture of benzyl4-(hydroxymethyl)-3,6-dihydropyridine-1(2H)-carboxylate (2.3 g, 9.3mmol) and PPh₃ (2.93 g, 11.2 mmol) in DCM at 0° C. was added CBr₄ (3.70g, 11.2 mmol) dropwise. The mixture was stirred at room temperatureuntil completion, then quenched with saturated NH₄Cl. The aqueous layerwas extracted with EtOAc (2×30 mL), the combined organic layers wereconcentrated under reduced pressure and the residue was purified bysilica gel column chromatography to give benzyl4-(bromomethyl)-3,6-dihydropyridine-1(2H)-carboxylate (2.1 g, 73% yield)as an oil. LCMS (ESI): m/z [M+H]⁺ calc'd for C₁₄H₁₆BrNO₂ 309.0; found310.0.

Step 4.

To a mixture of(S)-(4-bromothiazol-2-yl)((2S,5R)-3,6-diethoxy-5-isopropyl-2,5-dihydropyrazin-2-yl)methanol(1.37 g, 3.39 mmol) in DMF (20 mL) at 0° C. was added NaH (0.16 g, 6.78mmol) in portions. The mixture was stirred at 0° C. for 1 h, then benzyl4-(bromomethyl)-3,6-dihydropyridine-1(2H)-carboxylate (2.10 g, 6.78mmol) was added. The mixture was warmed to room temperature and stirredfor 2 h, then diluted with saturated NH₄Cl (100 mL), and the mixture wasextracted with EtOAc (2×30 mL). The combined organic layers wereconcentrated under reduced pressure and the residue was purified bysilica gel column chromatography to give benzyl4-(((S)-(4-bromothiazol-2-yl)((2S,5R)-3,6-diethoxy-5-isopropyl-2,5-dihydropyrazin-2-yl)methoxy)methyl)-3,6-dihydropyridine-1(2H)-carboxylate (1.75 g, 82% yield) as an oil. LCMS (ESI): m/z [M+H]⁺calc'd for C₂₉H₂₇BrN₄O₅S. 632.2; found 633.2.

Step 5.

To a mixture of benzyl4-(((S)-(4-bromothiazol-2-yl)((2S,5R)-3,6-diethoxy-5-isopropyl-2,5-dihydropyrazin-2-yl)methoxy)methyl)-3,6-dihydropyridine-1(2H)-carboxylate (1.75 g, 2.76 mmol) in THF (40 mL) and MECN (16 mL) at0° C. was added 0.02M HCl (35 mL, 0.7 mmol) dropwise. The mixture waswarmed to room temperature and stirred overnight, then quenched withsaturated NaHCO₃ and extracted with EtOAc (2×100 mL). The combinedorganic layers were concentrated under reduced pressure and the residuewas purified by silica gel column chromatography to give benzyl4-(((1S,2S)-2-amino-1-(4-bromothiazol-2-yl)-3-ethoxy-3-oxopropoxy)methyl)-3,6-dihydropyridine-1(2H)-carboxylate(1.15 g, 79% yield) as an oil. LCMS (ESI): m/z [M+H]⁺ calc'd forC₂₂H₂₆BrN₃O₅S. 523.1; found 524.2.

Step 6.

A mixture of benzyl4-(((1S,2S)-2-amino-1-(4-bromothiazol-2-yl)-3-ethoxy-3-oxopropoxy)methyl)-3,6-dihydropyridine-1(2H)-carboxylate(1.15 g, 2.19 mmol) and LiOH (0.21 g, 8.77 mmol) in THF (50 mL) and H₂O(50 mL) was stirred at room temperature for 1 h, then acidified to pH ˜4with 1M HCl. The mixture was then used directly in the next step withoutfurther purification. LCMS (ESI): m/z [M+H]⁺ calc'd for C₂₀H₂₂BrN₃O₅S.497.0; found 497.9 [for ⁸¹Br].

Step 7.

To the above mixture was added NaHCO₃ (0.97 g, 11.59 mmol) and (Boc)₂O(1.01 g, 4.63 mmol). The mixture was stirred at room temperatureovernight, then extracted with DCM (3×20 mL). The aqueous layer wasacidified to pH ˜4 with 1M HCl and extracted with EtOAc (3×30 mL). Thecombined organic layers were concentrated under reduced pressure to give(2S,3S)-3-((1-((benzyloxy)carbonyl)-1,2,3,6-tetrahydropyridin-4-yl)methoxy)-3-(4-bromothiazol-2-yl)-2-((tert-butoxycarbonyl)amino)propanoicacid (1.1 g, 79% yield) as an oil. LCMS (ESI): m/z [M+H]⁺ calc'd forC₂₅H₃₀BrN₃O₇S. 597.1; found 598.0 [for ⁸¹Br].

Step 8.

To a mixture of3-(1-ethyl-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)-2,2-dimethylpropyl(S)-hexahydropyridazine-3-carboxylate (1.5 g, 2.5 mmol) and(2S,3S)-3-((1-((benzyloxy)carbonyl)-1,2,3,6-tetrahydropyridin-4-yl)methoxy)-3-(4-bromothiazol-2-yl)-2-((tert-butoxycarbonyl)amino)propanoicacid (1.48 g, 2.5 mmol) in DMF was added DIPEA (3.21 g, 24.8 mmol) andHATU (1.89 g, 4.96 mmol) in portions. The mixture was stirred at roomtemperature for 2 h, then washed with H₂O (3×30 mL). The combinedaqueous layers were extracted with EtOAc (3×30 mL), the combined organiclayers were concentrated under reduced pressure and the residue waspurified by silica gel column chromatography to give3-(1-ethyl-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)-2,2-dimethylpropyl-(S)-1-((2S,3S)-3-((1-((benzyloxy)carbonyl)-1,2,3,6-tetrahydropyridin-4-yl)methoxy)-3-(4-bromothiazol-2-yl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylate(1.0 g, 34% yield) as an oil. LCMS (ESI): m/z [M+H]⁺ calc'd forC₅₉H₇₇BBrN₇O₁₁S. 1183.5; found 1184.3 [for ⁸¹Br].

Step 9.

To a mixture of3-(1-ethyl-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)-2,2-dimethylpropyl-(S)-1-((2S,3S)-3-((1-((benzyloxy)carbonyl)-1,2,3,6-tetrahydropyridin-4-yl)methoxy)-3-(4-bromothiazol-2-yl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylate(1.0 g, 0.85 mmol) and K₃PO₄ (0.45 g, 2.11 mmol) in toluene, dioxane andH₂O under an atmosphere of N₂ was added Pd(dtbpf)Cl₂ (0.11 g, 0.17 mmol)in portions. The mixture was heated to 60° C. and stirred for 2 h, thenwashed with H₂O (3×30 mL) and the combined aqueous layers extracted withEtOAc (3×30 mL). The combined organic layers were concentrated underreduced pressure and the residue was purified by silica gel columnchromatography to give benzyl4-((((6³S,3S,4S,Z)-4-((tert-butoxycarbonyl)amino)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-3-yl)oxy)methyl)-3,6-dihydropyridine-1(2H)-carboxylate(260 mg, 32% yield) as an oil. LCMS (ESI): m/z [M+H]⁺ calc'd forC₅₃H₆₅N₇O₉S. 975.5; found 976.6.

Step 10.

A mixture of benzyl4-((((6³S,3S,4S,Z)-4-((tert-butoxycarbonyl)amino)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-3-yl)oxy)methyl)-3,6-dihydropyridine-1(2H)-carboxylate(260 mg, 0.27 mmol) and Pd(OH)₂, 20% on carbon (0.26 g) in MeOH (3 mL)was stirred under an atmosphere of H₂ (balloon) at room temperature for1 h. The mixture was filtered through a pad of Celite pad and thefiltrate was concentrated under reduced pressure to give tert-butyl((6³S,3S,4S,Z)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-3-(piperidin-4-ylmethoxy)-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(230 mg, 72% yield) as an oil. LCMS (ESI): m/z [M+H]⁺ calc'd forC₄₅H₆₁N₇O₇S. 843.4; found 844.3.

Step 11.

To a mixture of tert-butyl((6³S,3S,4S,Z)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-3-(piperidin-4-ylmethoxy)-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(230 mg, 0.27 mmol) and AcOH (49 mg, 0.82 mmol) in MeOH was addedparaformaldehyde (49 mg, 1.6 mmol) and NaBH₃CN (86 mg, 1.36 mmol) inportions. The mixture was stirred at room temperature for 2 h, thendiluted with H₂O and extracted with EtOAc (3×20 mL). The combinedorganic layers were concentrated under reduced pressure and the residuewas purified by silica gel column chromatography to give tert-butyl((6³S,3S,4S,Z)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-3-((1-methylpiperidin-4-yl)methoxy)-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(280 mg, 81% yield) as a an oil. LCMS (ESI): m/z [M+H]⁺ calc'd forC₄₆H₆₃N₇O₇S. 857.5; found 858.4.

Step 12.

A mixture of tert-butyl ((6³S,3S,4S,Z)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-3-((1-methylpiperidin-4-yl)methoxy)-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(280 mg, 0.33 mmol) and HCl in 1,4-dioxane (3 mL) in 1,4-dioxane wasstirred at room temperature for 1 h. The mixture was concentrated underreduced pressure to give(6³S,3S,4S,Z)-4-amino-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-3-((1-methylpiperidin-4-yl)methoxy)-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-5,7-dioneas an oil. LCMS (ESI): m/z [M+H]⁺ calc'd for C₄₁H₅₅N₇O₅S. 757.4; found758.5.

Step 13.

To a mixture of(6³S,3S,4S,Z)-4-amino-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-3-((1-methylpiperidin-4-yl)methoxy)-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-5,7-dione(265 mg, 0.35 mmol) and (1S,2S)-2-methylcyclopropane-1-carboxylic acid(35 mg, 0.35 mmol) in DMF was added DIPEA (904 mg, 7.0 mmol) inportions. The mixture was stirred at room temperature for 2 h, thenwashed with H₂O (3×20 mL). The combined aqueous layers were extractedwith EtOAc (2×20 mL) and the combined organic layers were concentratedunder reduced pressure and the residue was purified by silica gel columnchromatography to give(1S,2S)—N-((6³S,3S,4S,Z)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-3-((1-methylpiperidin-4-yl)methoxy)-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2-methylcyclopropane-1-carboxamide(24 mg, 8% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₄₆H₆₁N₇O₆S. 839.4; found 840.4; ¹H NMR (300 MHz, DMSO-d₆) δ 8.83-8.64(m, 1H), 8.56-8.43 (m, 1H), 8.00-7.89 (m, 1H), 7.81-7.73 (m, 3H),7.63-7.53 (m, 2H), 5.93-5.85 (m, 1H), 5.20-5.12 (m, 1H), 4.90 (s, 1H),4.32-4.13 (m, 5H), 3.68-3.60 (m, 2H), 3.26-3.22 (m, 5H), 2.95-2.83 (m,1H), 2.79-2.72 (m, 3H), 2.49-2.46 (m, 1H), 2.20-2.10 (s, 3H), 2.09-2.01(m, 1H), 1.93-1.72 (m, 6H), 1.70-1.45 (m, 3H), 1.41-1.30 (m, 3H),1.21-0.99 (m, 7H), 0.98-0.87 (m, 8H), 0.60-0.50 (m, 1H), 0.35 (s, 3H).

Example A457. Synthesis of(1r,2R,3S)—N-((6³S,4S,Z)-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-1¹-(2-((tetrahydro-2H-pyran-4-yl)oxy)ethyl)-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2,3-dimethylcyclopropane-1-carboxamide

Step 1.

To a mixture of(S)-5-bromo-3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-2-(2-(1-methoxyethyl)pyridin-3-yl)-1H-indole(3.0 g, 4.6 mmol) and 2-((tetrahydro-2H-pyran-4-yl)oxy)ethyl4-methylbenzenesulfonate (2.06 g, 6.9 mmol) in DMF (20 mL) at 0° C.under an atmosphere of N₂ was added Cs₂CO₃ (3.73 g, 11.4 mmol) inportions. The mixture was heated to 65° C. and stirred overnight, thendiluted with H₂O (50 mL) and extracted with EtOAc (3×50 mL). Thecombined organic layers were washed with brine (3×50 mL), dried overanhydrous Na₂SO₄ and filtered. The filtrate was concentrated underreduced pressure and the residue was purified by silica gel columnchromatography to give(S)-5-bromo-3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-2-(2-(1-methoxyethyl)pyridin-3-yl)-1-(2-((tetrahydro-2H-pyran-4-yl)oxy)ethyl)-1H-indole(2.3 g, 64% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₄₄H₅₅BrN₂O₄Si, 784.3; found 785.2.

Step 2.

To a mixture of(S)-5-bromo-3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)-2-(2-(1-methoxyethyl)pyridin-3-yl)-1-(2-((tetrahydro-2H-pyran-4-yl)oxy)ethyl)-1H-indole(2.3 g, 2.9 mmol) in THF (20 mL) at 0° C. under an atmosphere of N₂ wasadded TBAF, 1M in THF (14.67 mL, 14.7 mmol) in portions. The mixture washeated to 45° C. and stirred for 6 h, then diluted with H₂O andextracted with EtOAc (3×30 mL). The combined organic layers were washedwith brine (3×30 mL), dried over anhydrous Na₂SO₄ and filtered. Thefiltrate was concentrated under reduced pressure and the residue waspurified by preparative-TLC to give(S)-3-(5-bromo-2-(2-(1-methoxyethyl)pyridin-3-yl)-1-(2-((tetrahydro-2H-pyran-4-yl)oxy)ethyl)-1H-indol-3-yl)-2,2-dimethylpropan-1-ol(690 mg, 43% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₂₈H₃₇BrN₂O₄ 546.2; found 546.9.

Step 3.

To a mixture of(S)-3-(5-bromo-2-(2-(1-methoxyethyl)pyridin-3-yl)-1-(2-((tetrahydro-2H-pyran-4-yl)oxy)ethyl)-1H-indol-3-yl)-2,2-dimethylpropan-1-ol(690 mg, 1.3 mmol) and (Bpin)₂ (643 mg, 2.5 mmol) in toluene (7 mL)under an atmosphere of Ar was added KOAc (372 mg, 3.8 mmol) andPd(dppf)Cl₂ (93 mg, 0.13 mmol) in portions. The mixture was heated to80° C. and stirred for 2.5 h, then diluted with H₂O (20 mL) andextracted with EtOAc (3×20 mL). The combined organic layers were washedwith brine (3×30 mL), dried over anhydrous Na₂SO₄ and filtered. Thefiltrate was concentrated under reduced pressure and the residue waspurified by preparative-TLC to give(S)-3-(2-(2-(1-methoxyethyl)pyridin-3-yl)-1-(2-((tetrahydro-2H-pyran-4-yl)oxy)ethyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)-2,2-dimethylpropan-1-ol(700 mg, 93% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₃₄H₄₉BN₂O₆ 592.4; found 593.1.

Step 4.

To a mixture of(S)-3-(2-(2-(1-methoxyethyl)pyridin-3-yl)-1-(2-((tetrahydro-2H-pyran-4-yl)oxy)ethyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)-2,2-dimethylpropan-1-ol(670 mg, 1.13 mmol) and Pd(DtBPF)Cl₂ (147 mg, 0.23 mmol) in toluene (3mL), 1,4-dioxane (1 mL) and H₂O (1 mL) under an atmosphere of Ar wasadded methyl(S)-1-((S)-3-(4-bromothiazol-2-yl)-2-((tert-butoxycarbonyl)amino)propanoyl)hexahydropyridazine-3-carboxylate(648 mg, 1.36 mmol) and K₃PO₄ (720 mg, 3.39 mmol) in portions. Themixture was heated to 60° C. and stirred for 3 h, then diluted with H₂O(10 mL) and extracted with EtOAc (3×10 mL). The combined organic layerswere washed with brine (3×10 mL), dried over anhydrous Na₂SO₄ andfiltered. The filtrate was concentrated under reduced pressure and theresidue was purified by preparative-TLC to give methyl(S)-1-((S)-2-((tert-butoxycarbonyl)amino)-3-(4-(3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2-((tetrahydro-2H-pyran-4-yl)oxy)ethyl)-1H-indol-5-yl)thiazol-2-yl)propanoyl)hexahydropyridazine-3-carboxylate(680 mg, 70% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₄₅H₆₂N₆O₉S. 862.4; found 863.1.

Step 5.

To a mixture of methyl(S)-1-((S)-2-((tert-butoxycarbonyl)amino)-3-(4-(3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2-((tetrahydro-2H-pyran-4-yl)oxy)ethyl)-1H-indol-5-yl)thiazol-2-yl)propanoyl)hexahydropyridazine-3-carboxylate(300 mg, 0.35 mmol) in THF (6 mL) at 0° C. under an atmosphere of N₂ wasadded 1M LiOH (1.74 mL, 1.74 mmol) in portions. When the reaction wasdeemed complete by LCMS the mixture was acidified to pH ˜6 with 1M HCl,then extracted with EtOAc (3×50 mL). The combined organic layers werewashed with brine (2×20 mL), dried over anhydrous Na₂SO₄, filtered andthe filtrate was concentrated under reduced pressure to give(S)-1-((S)-2-((tert-butoxycarbonyl)amino)-3-(4-(3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2-((tetrahydro-2H-pyran-4-yl)oxy)ethyl)-1H-indol-5-yl)thiazol-2-yl)propanoyl)hexahydropyridazine-3-carboxylicacid (280 mg) as a solid, which was used directly in the next stepwithout further purification. LCMS (ESI): m/z [M+H]⁺ calc'd forC₄₄H₆₀N₆O₉S. 848.4; found 849.4.

Step 6.

To a mixture of(S)-1-((S)-2-((tert-butoxycarbonyl)amino)-3-(4-(3-(3-hydroxy-2,2-dimethylpropyl)-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2-((tetrahydro-2H-pyran-4-yl)oxy)ethyl)-1H-indol-5-yl)thiazol-2-yl)propanoyl)hexahydropyridazine-3-carboxylicacid (300 mg, 0.35 mmol) and DIPEA (457 mg, 3.5 mmol) in DCM (50 mL) at0° C. under an atmosphere of N₂ was added HOBT (477 mg, 3.5 mmol) andEDCl (2.03 g, 10.6 mmol) in portions. The mixture was warmed to roomtemperature and stirred overnight, then washed with H₂O (50 mL) and theaqueous layer was extracted with DCM (3×50 mL). The combined organiclayers were washed with brine (3×100 mL), dried over anhydrous Na₂SO₄and filtered. The filtrate was concentrated under reduced pressure andthe residue was purified by preparative-TLC to give tert-butyl((6³S,4S,Z)-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-1¹-(2-((tetrahydro-2H-pyran-4-yl)oxy)ethyl)-6¹,6²,6³,6⁴,65,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(200 mg, 68% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₄₄H₅₈N₆O₈S. 830.4; found 831.3.

Step 7.

To a mixture of tert-butyl((6³S,4S,Z)-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-1¹-(2-((tetrahydro-2H-pyran-4-yl)oxy)ethyl)-6¹,6²,6³,6⁴,65,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(90 mg, 0.11 mmol) in DCM (1 mL) at 0° C. under an atmosphere of N₂ wasadded TFA (1 mL) in portions. The mixture was stirred at 0° C. for 1.5h, then concentrated under reduced pressure to give(6³S,4S,Z)-4-amino-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-1¹-(2-((tetrahydro-2H-pyran-4-yl)oxy)ethyl)-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-5,7-dioneTFA salt (80 mg) as an oil, that was used directly in the next stepwithout further purification. LCMS (ESI): m/z [M+H]⁺ calc'd forC₃₉H₅₀N₆O₆S. 730.4; found 731.4.

Step 8.

To a mixture of(6³S,4S,Z)-4-amino-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-1¹-(2-((tetrahydro-2H-pyran-4-yl)oxy)ethyl)-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-5,7-dioneTFA salt (90 mg, 0.12 mmol) and(1r,2R,3S)-2,3-dimethylcyclopropane-1-carboxylic acid (17 mg, 0.15 mmol)at 0° C. under an atmosphere of N₂ was added DIPEA (318 mg, 2.5 mmol)and HATU (56 mg, 0.15 mmol) in portions. The mixture was stirred at 0°C. for 1 h, then washed with H₂O (1 mL) and extracted with EtOAc (3×30mL). The combined organic layers were washed with brine (30 mL), driedover anhydrous Na₂SO₄ and filtered. The filtrate was concentrated underreduced pressure and the residue was purified by preparative-HPLC togive(1r,2R,3S)—N-((6³S,4S,Z)-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-1¹-(2-((tetrahydro-2H-pyran-4-yl)oxy)ethyl)-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2,3-dimethylcyclopropane-1-carboxamide(17 mg, 17% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₄₅H₅₈N₆O₇S. 826.4; found 827.6; ¹H NMR (400 MHz, DMSO-d₆) δ 8.74 (dd,J=4.7, 1.8 Hz, 1H), 8.48 (d, J=1.6 Hz, 1H), 8.39 (d, J=9.0 Hz, 1H), 7.81(s, 1H), 7.74 (ddd, J=10.5, 8.2, 1.8 Hz, 2H), 7.61 (d, J=8.7 Hz, 1H),7.51 (dd, J=7.7, 4.7 Hz, 1H), 5.56 (t, J=9.0 Hz, 1H), 5.07 (d, J=12.1Hz, 1H), 4.46 (dt, J=14.9, 5.3 Hz, 1H), 4.32-4.09 (m, 4H), 3.62-3.47 (m,4H), 3.34-3.30 (m, 1H), 3.30-3.28 (m, 1H), 3.26 (s, 3H), 3.21-3.04 (m,4H), 2.94 (d, J=14.3 Hz, 1H), 2.83-2.68 (m, 1H), 2.46-2.43 (m, 1H),2.16-2.05 (m, 1H), 1.76 (d, J=20.9 Hz, 2H), 1.51 (d, J=13.5 Hz, 3H),1.36 (d, J=6.1 Hz, 3H), 1.30-1.09 (m, 3H), 1.13-0.98 (m, 8H), 0.88 (s,3H), 0.32 (s, 3H).

Example A459. Synthesis of(2S)—N-((6³S,3S,4S,Z)-3-ethoxy-1¹-(2-isopropoxyethyl)-1²-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2-(methoxymethyl)azetidine-1-carboxamide

Step 1.

A mixture of(6³S,3S,4S,Z)-4-amino-3-ethoxy-1¹-(2-isopropoxyethyl)-1²-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-5,7-dione(170 mg, 0.21 mmol) in THF (6 mL) at 0° C. under an atmosphere of N₂ wasadded TEA (62 mg, 0.62 mmol) and 4-nitrophenyl chloroformate (62 mg,0.31 mmol) in portions. The mixture was warmed to room temperature andstirred for 4 h, then concentrated under reduced pressure to give4-nitrophenyl((6³S,3S,4S,Z)-3-ethoxy-1¹-(2-isopropoxyethyl)-12-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(200 mg) as a solid, which was used directly in the next step withoutfurther purification.

Step 2.

To a mixture of 4-nitrophenyl((6³S,3S,4S,Z)-3-ethoxy-1¹-(2-isopropoxyethyl)-12-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(150 mg, 0.15 mmol) in MeCN (8 mL) at 0° C. under an atmosphere of N₂was added DIPEA (78 mg, 0.60 mmol) and (2S)-2-(methoxymethyl)azetidineHCl salt (46 mg, 0.45 mmol) in portions. The mixture was warmed to roomtemperature and stirred overnight, then the mixture was diluted withbrine (10 mL) and extracted with EtOAc (3×20 mL). The combined organiclayers were washed with brine (3×20 mL), dried over anhydrous Na₂SO₄ andfiltered. The filtrate was concentrated under reduced pressure and theresidue was purified by preparative-HPLC to give(2S)—N-((6³S,3S,4S,Z)-3-ethoxy-1¹-(2-isopropoxyethyl)-1²-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2-(methoxymethyl)azetidine-1-carboxamide(56 mg, 39% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₅₀H₇₁N₉O₈S. 957.5; found 958.4; ¹H NMR (400 MHz, DMSO-d₆) δ 8.47 (dd,J=14.9, 2.2 Hz, 2H), 7.91 (s, 1H), 7.74 (dd, J=8.6, 1.6 Hz, 1H), 7.59(d, J=8.7 Hz, 1H), 7.21 (s, 1H), 6.83 (d, J=10.2 Hz, 1H), 5.51 (d,J=10.2 Hz, 1H), 5.16 (d, J=12.0 Hz, 1H), 4.90 (s, 1H), 4.37 (s, 2H),4.27-4.00 (m, 4H), 3.71 (q, J=8.4 Hz, 1H), 3.65-3.54 (m, 5H), 3.50 (q,J=9.4, 8.4 Hz, 2H), 3.43 (s, 3H), 3.40 (s, 2H), 3.30 (s, 1H), 3.29-3.20(m, 4H), 3.13 (s, 3H), 2.90-2.72 (m, 2H), 2.67 (s, 1H), 2.43 (s, 2H),2.23 (s, 4H), 2.05 (d, J=12.0 Hz, 1H), 1.82 (dd, J=23.8, 13.9 Hz, 3H),1.52 (d, J=12.1 Hz, 1H), 1.33 (d, J=6.1 Hz, 3H), 1.13 (t, J=7.0 Hz, 3H),0.93-0.84 (m, 6H), 0.81 (d, J=6.1 Hz, 3H), 0.44 (s, 3H).

Example A460. Synthesis of(2R)—N-((6³S,3S,4S,Z)-3-ethoxy-1¹-(2-isopropoxyethyl)-1²-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2-methylazetidine-1-carboxamide

Step 1.

To a mixture of 4-nitrophenyl((6³S,3S,4S,Z)-3-ethoxy-1¹-(2-isopropoxyethyl)-1²-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(150 mg, 0.15 mmol) in MECN (8 mL) at 0° C. under an atmosphere of N₂was added DIPEA (78 mg, 0.60 mmol) and (2R)-2-methylazetidine (32 mg,0.45 mmol) in portions. The mixture was warmed to room temperature andstirred overnight, then diluted with brine (10 mL) and extracted withEtOAc (3×20 mL). The combined organic layers were washed with brine(3×20 mL), dried over anhydrous Na₂SO₄ and filtered. The filtrate wasconcentrated under reduced pressure and the residue was purified bypreparative-HPLC to give(2R)—N-((6³S,3S,4S,Z)-3-ethoxy-1¹-(2-isopropoxyethyl)-1²-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2-methylazetidine-1-carboxamide(57 mg, 41% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₄₉H₆₉N₉O₇S. 927.5; found 928.4; ¹H NMR (400 MHz, DMSO-d₆) δ 8.53-8.40(m, 2H), 7.93 (s, 1H), 7.78-7.69 (m, 1H), 7.59 (d, J=8.6 Hz, 1H), 7.21(s, 1H), 5.56 (d, J=10.2 Hz, 1H), 5.21 (d, J=12.1 Hz, 1H), 5.12 (d,J=10.2 Hz, 1H), 4.91 (s, 1H), 4.35 (d, J=13.9 Hz, 1H), 4.27-4.12 (m,4H), 4.08 (d, J=6.1 Hz, 1H), 3.75 (t, J=7.4 Hz, 2H), 3.65-3.46 (m, 4H),3.30 (s, 2H), 3.29 (s, 1H), 3.26 (d, J=5.9 Hz, 5H), 3.13 (s, 3H), 2.79(d, J=13.5 Hz, 2H), 2.60 (s, 2H), 2.45 (p, J=1.9 Hz, 4H), 2.22 (s, 3H),2.06 (d, J=12.1 Hz, 1H), 1.82 (s, 3H), 1.54-1.49 (m, 1H), 1.34 (dd,J=14.9, 6.2 Hz, 6H), 1.13 (t, J=7.0 Hz, 3H), 0.87 (t, J=6.8 Hz, 6H),0.81 (d, J=6.0 Hz, 3H), 0.44 (s, 3H).

Example A476. Synthesis of(1R,2R,3S)—N-((3′S,3′S,4′S,Z)-3′-ethoxy-1′-(2-isopropoxyethyl)-2′-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-5′,7′-dioxospiro[cyclopropane-1,10′-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphan]-4′-yl)-2,3-dimethylcyclopropane-1-carboxamideand(1R,2R,3S)—N-((3′S,3′S,4′S,Z)-3′-ethoxy-1′-(2-isopropoxyethyl)-2′-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-5′,7′-dioxospiro[cyclopropane-1,10′-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphan]-4′-yl)-2,3-dimethylcyclopropane-1-carboxamide

Step 1.

To a mixture of5-bromo-3-((1-(((tert-butyldiphenylsilyl)oxy)methyl)cyclopropyl)methyl)-2-iodo-1H-indole(6.1 g, 9.5 mmol) and benzyl(S)-4-(6-(1-methoxyethyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-3-yl)piperazine-1-carboxylate(4.56 g, 9.5 mmol) in 1,4-dioxane (120 mL) and H₂O (20 mL) under anatmosphere of N₂ was added Pd(dppf)Cl₂ (1.04 g, 1.4 mmol) and K₃PO₄(4.02 g, 18.9 mmol) in portions. The mixture was heated to 65° C. andstirred for 3 h, then extracted with EtOAc (3×100 mL). The combinedorganic layers were washed with brine (3×100 mL), dried over anhydrousNa₂SO₄ and filtered. The filtrate was concentrated under reducedpressure and the residue was purified by silica gel columnchromatography to give benzyl(S)-4-(5-(5-bromo-3-((1-(((tert-butyldiphenylsilyl)oxy)methyl)cyclopropyl)methyl)-1H-indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylate(4.7 g, 57% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₄₉H₅₅BrN₄O₄Si, 870.3; found 871.5.

Step 2.

To a mixture of benzyl(S)-4-(5-(5-bromo-3-((1-(((tert-butyldiphenylsilyl)oxy)methyl)cyclopropyl)methyl)-1H-indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylate(4.6 g, 5.3 mmol) in DMF (50 mL) at 0° C. was added 2-isopropoxyethyl4-methylbenzenesulfonate (2.04 g, 7.9 mmol) and Cs₂CO₃ (5.16 g, 15.8mmol). The mixture was heated to 50° C. and stirred for 2 h, thendiluted with H₂O and extracted with EtOAc (2×100 mL). The combinedorganic layers were washed with brine (3×100 mL), dried over anhydrousNa₂SO₄ and filtered. The filtrate was concentrated under reducedpressure and the residue was purified by reverse-phase silica gel columnchromatography to give benzyl(S)-4-(5-(5-bromo-3-((1-(((tert-butyldiphenylsilyl)oxy)methyl)cyclopropyl)methyl)-1-(2-isopropoxyethyl)-1H-indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylate(4.5 g, 89% yield) as an oil. LCMS (ESI): m/z [M+H]⁺ calc'd forC₅₄H₆₅BrN₄O₅Si, 956.4; found 957.2.

Step 3.

To a mixture of benzyl(S)-4-(5-(5-bromo-3-((1-(((tert-butyldiphenylsilyl)oxy)methyl)cyclopropyl)methyl)-1-(2-isopropoxyethyl)-1H-indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylate(3.2 g, 3.3 mmol) in THF (32 mL) at 0° C. was added TBAF (13.1 g, 50.1mmol). The mixture was warmed to room temperature and stirred for 16 h,then diluted with H₂O and extracted with EtOAc (2×100 mL). The combinedorganic layers were washed with brine (3×100 mL), dried over anhydrousNa₂SO₄ and filtered. The filtrate was concentrated under reducedpressure and the residue was purified by reverse-phase silica gel columnchromatography to give benzyl(S)-4-(5-(5-bromo-3-((1-(hydroxymethyl)cyclopropyl)methyl)-1-(2-isopropoxyethyl)-1H-indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylate(1.84 g, 77% yield) as an oil. LCMS (ESI): m/z [M+H]⁺ calc'd forC₃₈H₄₇BrN₄O₅ 718.3; found 719.2.

Step 4.

To a mixture of benzyl(S)-4-(5-(5-bromo-3-((1-(hydroxymethyl)cyclopropyl)methyl)-1-(2-isopropoxyethyl)-1H-indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylate(1.7 g, 2.4 mmol) in toluene (20 mL) at 0° C. under an atmosphere of Arwas added 4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi(1,3,2-dioxaborolane)(1.80 g, 7.1 mmol), KOAc (580 mg, 5.9 mmol) and Pd(dppf)Cl₂ (346 mg,0.47 mmol). The mixture was heated to 90° C. and stirred for 3 h, thendiluted with H₂O and extracted with EtOAc (2×50 mL). The combinedorganic layers were washed with brine (2×50 mL), dried over anhydrousNa₂SO₄ and filtered. The filtrate was concentrated under reducedpressure and the residue was purified by reverse-phase silica gel columnchromatography to give benzyl(S)-4-(5-(3-((1-(hydroxymethyl)cyclopropyl)methyl)-1-(2-isopropoxyethyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylate(1.4 g, 77% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₄₄H₅₉BN₄O₇ 766.5; found 767.4.

Step 5.

To a mixture of benzyl(S)-4-(5-(3-((1-(hydroxymethyl)cyclopropyl)methyl)-1-(2-isopropoxyethyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylate(1.4 g, 1.8 mmol) in DCM (20 mL) at 0° C. was added(S)-1,2-bis(tert-butoxycarbonyl)hexahydropyridazine-3-carboxylic acid(664 mg, 2.0 mmol), DCC (490 mg, 2.4 mmol) and DMAP (45 mg, 0.37 mmol).The mixture was warmed to room temperature and stirred for 16 h, thendiluted with H₂O and extracted with EtOAc (2×100 mL). The combinedorganic layers were washed with brine (2×100 mL), dried over anhydrousNa₂SO₄ and filtered. The filtrate was concentrated under reducedpressure and the residue was purified by reverse-phase silica gel columnchromatography to give3-((1-((2-(5-(4-((benzyloxy)carbonyl)piperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2-isopropoxyethyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)methyl)cyclopropyl)methyl)1,2-di-tert-butyl (S)-tetrahydropyridazine-1,2,3-tricarboxylate (1.2 g,61% yield) as an oil. LCMS (ESI): m/z [M+H]⁺ calc'd for C₅₉H₈₃BN₆O₁₂1078.6; found 1079.5.

Step 6.

A mixture of3-((1-((2-(5-(4-((benzyloxy)carbonyl)piperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2-isopropoxyethyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)methyl)cyclopropyl)methyl)1,2-di-tert-butyl (S)-tetrahydropyridazine-1,2,3-tricarboxylate (1.2 g,1.1 mmol) in HCl in 1,4-dioxane (15 mL) at 0° C. was stirred for 2 h,then concentrated under reduced pressure to give(1-((2-(5-(4-((benzyloxy)carbonyl)piperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2-isopropoxyethyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)methyl)cyclopropyl)methyl(S)-hexahydropyridazine-3-carboxylate (1.25 g), which was used directlyin the next step without further purification. LCMS (ESI): m/z [M+H]⁺calc'd for C₄₉H₆₇BN₆O₈ 878.5; found 879.4.

Step 7.

To a mixture of(1-((2-(5-(4-((benzyloxy)carbonyl)piperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2-isopropoxyethyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)methyl)cyclopropyl)methyl(S)-hexahydropyridazine-3-carboxylate (1.0 g, 1.1 mmol) in DMF (15 mL)at 0° C. was added DIPEA (1.47 g, 11.4 mmol) and(2S,3S)-3-(4-bromothiazol-2-yl)-2-((tert-butoxycarbonyl)amino)-3-ethoxypropanoicacid (675 mg, 1.7 mmol), followed by HATU (865 mg, 2.3 mmol). Themixture was stirred at 0° C. for 2 h, then diluted with H₂O andextracted with EtOAc (2×100 mL). The combined organic layers were washedwith brine (2×100 mL), dried over anhydrous Na₂SO₄ and filtered. Thefiltrate was concentrated under reduced pressure and the residue waspurified by preparative-TLC to give(1-((2-(5-(4-((benzyloxy)carbonyl)piperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2-isopropoxyethyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)methyl)cyclopropyl)methyl(S)-1-((2S,3S)-3-(4-bromothiazol-2-yl)-2-((tert-butoxycarbonyl)amino)-3-ethoxypropanoyl)hexahydropyridazine-3-carboxylate(730 mg, 51% yield) as an oil. LCMS (ESI): m/z [M+H]⁺ calc'd forC₆₂H₈₄BBrN₈O₁₂S. 1254.5; found 1255.4.

Step 8.

To a mixture of(1-((2-(5-(4-((benzyloxy)carbonyl)piperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2-isopropoxyethyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)methyl)cyclopropyl)methyl(S)-1-((2S,3S)-3-(4-bromothiazol-2-yl)-2-((tert-butoxycarbonyl)amino)-3-ethoxypropanoyl)hexahydropyridazine-3-carboxylate(700 mg, 0.56 mmol) in toluene (12 mL), 1,4-dioxane (4 mL) and H₂O (4mL) at 0° C. under an atmosphere of Ar was added XPhos (53 mg, 0.11mmol), K₃PO₄ (296 mg, 1.39 mmol) and XPho-Pd-G3 (47 mg, 0.06 mmol). Themixture was heated to 65° C. and stirred for 2 h, then diluted with H₂Oand extracted with EtOAc (2×50 mL). The combined organic layers werewashed with brine (2×50 mL), dried over anhydrous Na₂SO₄ and filtered.The filtrate was concentrated under reduced pressure and the residue waspurified by preparative-TLC to give benzyl4-(5-((3'S,3'S,4'S,Z)-4′-((tert-butoxycarbonyl)amino)-3′-ethoxy-1′-(2-isopropoxyethyl)-5′,7′-dioxospiro[cyclopropane-1,10′-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphan]-2′-yl)-6-((S)-1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylate(160 mg, 27% yield) as an oil. LCMS (ESI): m/z [M+H]⁺ calc'd forC₅₆H₇₂N₈O₁₀S. 1048.5; found 1049.3.

Step 9.

To a mixture of benzyl4-(5-((3'S,3'S,4'S,Z)-4′-((tert-butoxycarbonyl)amino)-3′-ethoxy-1′-(2-isopropoxyethyl)-5′,7′-dioxospiro[cyclopropane-1,10′-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphan]-2′-yl)-6-((S)-1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylate(160 mg, 0.15 mmol) in MeOH (3 mL) at 0° C. was added methoxymethanolamine (34 mg, 0.76 mmol) and Pd(OH)₂/C (171 mg, 1.2 mmol). The mixturewas placed under an atmosphere of H₂, heated to 35° C. and stirred for 4h, then filtered, and the filter cake was washed with MeOH (2×50 mL).The filtrate was concentrated under reduced pressure to give tert-butyl((3′S,3′S,4′S,Z)-3′-ethoxy-1′-(2-isopropoxyethyl)-2′-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-5′,7′-dioxospiro[cyclopropane-1,10′-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphan]-4′-yl)carbamate(100 mg), which was used directly in the next step without furtherpurification. LCMS (ESI): m/z [M+H]⁺ calc'd for C₄₉H₆₈N₈O₈S 928.5; found929.4.

Step 10.

To a mixture of tert-butyl((3′S,3′S,4′S,Z)-3′-ethoxy-1′-(2-isopropoxyethyl)-2′-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-5′,7′-dioxospiro[cyclopropane-1,10′-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphan]-4′-yl)carbamate(100 mg, 0.11 mmol) in DCM (1 mL) at 0° C. was added HCl in 1,4-dioxane(1 mL). The mixture was stirred at 0° C. for 2 h, then concentratedunder reduced pressure to give(3′S,3′S,4′S,Z)-4′-amino-3′-ethoxy-1′-(2-isopropoxyethyl)-2′-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)spiro[cyclopropane-1,10′-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane]-5′,7′-dione(110 mg), which was used directly in the next step without furtherpurification. LCMS (ESI): m/z [M+H]⁺ calc'd for C₄₄H₆₀N₈O₆S. 828.4;found 829.4.

Step 11.

To a mixture of(3′S,3′S,4′S,Z)-4′-amino-3′-ethoxy-1′-(2-isopropoxyethyl)-2′-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)spiro[cyclopropane-1,10′-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane]-5′,7′-dione(100 mg, 0.12 mmol) in DMF (2 mL) at 0° C. was added DIPEA (78 mg, 0.61mmol), (1R,2R,3S)-2,3-dimethylcyclopropane-1-carboxylic acid (21 mg,0.18 mmol) and HATU (92 mg, 0.24 mmol). The mixture was stirred at 0° C.for 2 h, then diluted with H₂O and extracted with EtOAc (2×30 mL). Thecombined organic layers were washed with brine (2×30 mL), dried overanhydrous Na₂SO₄ and filtered. The filtrate was concentrated underreduced pressure and the residue was purified by preparative-HPLC togive(1R,2R,3S)—N-((3′S,3′S,4′S,Z)-3′-ethoxy-1′-(2-isopropoxyethyl)-2′-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-5′,7′-dioxospiro[cyclopropane-1,10′-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphan]-4′-yl)-2,3-dimethylcyclopropane-1-carboxamide(7 mg, 6% yield) and(1R,2R,3S)—N-((3′S,3′S,4′S,Z)-3′-ethoxy-1′-(2-isopropoxyethyl)-2′-(2-((S)-1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-5′,7′-dioxospiro[cyclopropane-1,10′-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphan]-4′-yl)-2,3-dimethylcyclopropane-1-carboxamide(13 mg, 12% yield), both as solids. LCMS (ESI): m/z [M+H]⁺ calc'd forC₅₀H₆₈N₈O₇S. 924.5; found 925.5; 1H NMR (400 MHz, DMSO-d₆) δ 8.50 (d,J=1.5 Hz, 1H), 8.42 (d, J=3.0 Hz, 1H), 7.91 (s, 1H), 7.71 (d, J=8.6 Hz,1H), 7.64 (d, J=10.1 Hz, 1H), 7.58 (d, J=8.6 Hz, 1H), 7.25 (d, J=2.8 Hz,1H), 6.55 (s, 1H), 5.77 (d, J=10.1 Hz, 1H), 5.04 (d, J=12.3 Hz, 1H),4.92 (s, 1H), 4.37-4.28 (m, 1H), 4.22 (d, J=10.9 Hz, 1H), 4.18-4.08 (m,3H), 3.98 (dd, J=12.2, 6.1 Hz, 3H), 3.65-3.58 (m, 6H), 3.36-3.20 (m,6H), 3.17 (d, J=5.2 Hz, 2H), 2.98 (s, 3H), 2.45 (s, 6H), 2.39-2.30 (m,3H), 2.10-1.89 (m, 3H), 1.86-1.66 (m, 3H), 1.51 (d, J=3.8 Hz, 2H), 1.36(d, J=6.2 Hz, 3H), 1.24 (s, 1H), 1.16 (t, J=7.0 Hz, 5H), 1.08 (d, J=5.5Hz, 3H), 1.04 (d, J=5.5 Hz, 3H), 0.91 (d, J=6.1 Hz, 3H), 0.86 (d, J=6.1Hz, 4H) and LCMS (ESI): m/z [M+H]⁺ calc'd for C₅₀H₆₈N₈O₇S. 924.5; found925.5; ¹H NMR (400 MHz, DMSO-d₆) δ 8.52 (s, 1H), 8.47 (d, J=3.1 Hz, 1H),7.76-7.71 (m, 1H), 7.67 (d, J=10.2 Hz, 1H), 7.57 (d, J=8.5 Hz, 1H), 7.37(d, J=3.2 Hz, 1H), 5.76 (d, J=10.1 Hz, 1H), 5.03 (d, J=12.3 Hz, 1H),4.92 (s, 1H), 4.15 (dd, J=27.1, 12.3 Hz, 1H), 3.83 (d, J=6.2 Hz, 3H),3.54 (dd, J=17.3, 6.9 Hz, 1H), 3.44 (d, J=6.7 Hz, 3H), 3.34-3.26 (m,9H), 2.92 (s, 3H), 2.47-2.41 (m, 7H), 2.37-2.31 (m, 3H), 2.23 (s, 1H),1.94-1.77 (m, 3H), 1.55 (s, 2H), 1.24 (d, J=6.4 Hz, 4H), 1.15 (t, J=6.9Hz, 5H), 1.09 (d, J=5.7 Hz, 3H), 1.04 (d, J=5.7 Hz, 3H), 0.95 (d, J=6.1Hz, 3H), 0.86 (d, J=6.1 Hz, 3H), 0.50-0.42 (m, 1H), 0.33 (s, 2H),0.14-0.06 (m, 1H).

Example A483. Synthesis of(1r,2R,3S)—N-((6³S,3S,4S,Z)-1²-(5-((1R,5S,7s)-9-cyclopropyl-3-oxa-9-azabicyclo[3.3.1]nonan-7-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-3-ethoxy-1¹-ethyl-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2,3-dimethylcyclopropane-1-carboxamide

Step 1.

To a mixture of tert-butyl((6³S,3S,4S,Z)-1²-(5-((1R,5S,7s)-3-oxa-9-azabicyclo[3.3.1]nonan-7-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-3-ethoxy-1¹-ethyl-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(50% purity; 500 mg, 0.28 mmol) and(1-ethoxycyclopropoxy)trimethylsilane (968 mg, 5.6 mmol) in MeOH (2 mL)at 0° C. was added AcOH (83 mg, 1.39 mmol) and NaBH₃CN (87 mg, 1.39mmol) in portions. The mixture was warmed to 60° C. and stirred for 2 h,then concentrated under reduced pressure. The residue purified by silicagel column chromatography to give tert-butyl((6³S,3S,4S,Z)-1²-(5-((1R,5S,7s)-9-cyclopropyl-3-oxa-9-azabicyclo[3.3.1]nonan-7-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-3-ethoxy-1¹-ethyl-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(80 mg, 30% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₅₁H₆₉N₇O₈S. 939.5; found 940.6.

Step 2.

A mixture of tert-butyl ((6³S,3S,4S,Z)-1²-(5-((1R,5S,7s)-9-cyclopropyl-3-oxa-9-azabicyclo[3.3.1]nonan-7-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-3-ethoxy-1¹-ethyl-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(80 mg, 0.09 mmol) and TFA (0.4 mL) in DCM (2 mL) at 0° C. was stirredfor 1 h, then concentrated under reduced pressure to give(6³S,3S,4S,Z)-4-amino-1²-(5-((1R,5S,7s)-9-cyclopropyl-3-oxa-9-azabicyclo[3.3.1]nonan-7-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-3-ethoxy-1¹-ethyl-10,10-dimethyl-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-5,7-dione as a TFA salt (180 mg) as an oil.LCMS (ESI): m/z [M+H]⁺ calc'd for C₄₆H₆₁N₇O₆S. 839.4; found 840.4.

Step 3.

To a mixture of(6³S,3S,4S,Z)-4-amino-1²-(5-((1R,5S,7s)-9-cyclopropyl-3-oxa-9-azabicyclo[3.3.1]nonan-7-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-3-ethoxy-1¹-ethyl-10,10-dimethyl-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-5,7-dione(180 mg, 0.21 mmol) and (1R,2R,3S)-2,3-dimethylcyclopropane-1-carboxylicacid (49 mg, 0.43 mmol) in DMF (3 mL) at 0° C. was added DIPEA (277 mg,2.14 mmol) and COMU (184 mg, 0.43 mmol) in portions. The mixture waswarmed to room temperature and stirred for 2 h. The residue was purifiedby preparative-HPLC to give(1r,2R,3S)—N-((6³S,3S,4S,Z)-1²-(5-((1R,5S,7s)-9-cyclopropyl-3-oxa-9-azabicyclo[3.3.1]nonan-7-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-3-ethoxy-1¹-ethyl-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2,3-dimethylcyclopropane-1-carboxamide(23 mg, 11% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₅₂H₆₉N₇O₇S. 935.5; found 936.4; ¹H NMR (300 MHz, DMSO-d₆) δ 8.74 (d,J=2.2 Hz, 1H), 8.56 (s, 1H), 7.98 (s, 1H), 7.81 (d, J=8.6 Hz, 1H),7.74-7.56 (m, 3H), 5.94 (d, 0=9.7 Hz, 1H), 5.23 (d, J=12.4 Hz, 1H), 5.00(s, 1H), 4.42-4.08 (m, 5H), 3.92 (d, J=10.9 Hz, 2H), 3.90-3.65 (m, 6H)3.57-3.46 (m, 2H), 3.26 (s, 4H), 3.02 (d, J=10.1 Hz, 2H), 2.89 (m, 2H),2.67 (m, 1H), 2.35 (s, 2H), 2.13 (d, J=10.5 Hz, 1H), 1.85 (d, J=13.6 Hz,3H), 1.59 (s, 2H), 1.43 (d, J=6.1 Hz, 3H), 1.42-1.30 (m, 2H), 1.28-1.08(m, 12H), 1.10-0.98 (m, 7H), 0.55 (d, J=6.0 Hz, 2H), 0.45 (s, 2H), 0.39(s, 2H).

Example A484. Synthesis of(6³S,3S,4S,Z)-4-amino-3-ethoxy-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-((1R,5S,7s)-9-(oxetan-3-yl)-3-oxa-9-azabicyclo[3.3.1]nonan-7-yl)pyridin-3-yl)-10,10-dimethyl-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-5,7-dione

Step 1.

A mixture of3-(3-(5-bromo-1-ethyl-2-(2-((S)-1-methoxyethyl)-5-(9-((2,2,2-trichloroethoxy)carbonyl)-3-oxa-9-azabicyclo[3.3.1]non-6-en-7-yl)pyridin-3-yl)-1H-indol-3-yl)-2,2-dimethylpropyl)-1,2-di-tert-butyl-(3S)-tetrahydropyridazine-1,2,3-tricarboxylate(6.57 g, 6.22 mmol) and CsF (4.72 g, 31.1 mmol) in DMF (50 mL) washeated to 80° C. and stirred for 2 h, then diluted with H₂O (3×200 mL)and extracted with EtOAc (200 mL). The organic layer was concentratedunder reduced pressure to give3-(3-(2-(5-(3-oxa-9-azabicyclo[3.3.1]non-6-en-7-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-5-bromo-1-ethyl-1H-indol-3-yl)-2,2-dimethylpropyl)-1,2-di-tert-butyl-(3S)-tetrahydropyridazine-1,2,3-tricarboxylate(5.83 g) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd for C₄₅H₆₂BrN₅O₈879.4 & 881.4; found 880.1 & 882.1.

Step 2.

To a mixture of3-(3-(2-(5-(3-oxa-9-azabicyclo[3.3.1]non-6-en-7-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-5-bromo-1-ethyl-1H-indol-3-yl)-2,2-dimethylpropyl)-1,2-di-tert-butyl-(3S)-tetrahydropyridazine-1,2,3-tricarboxylate(5.83 g, 6.62 mmol) and NaHCO₃ (2.78 g, 33.1 mmol) in H₂O (20 mL) andTHF (20 mL) at 0° C. was added 9H-fluoren-9-ylmethyl chloroformate (2.57g, 9.93 mmol) in portions. The mixture was warmed to room temperatureand stirred overnight, then extracted with EtOAc (3×50 mL). The combinedorganic layers were dried over anhydrous Na₂SO₄, filtered, the filtratewas concentrated under reduced pressure and the residue was purified bysilica gel column chromatography to give3-(3-(2-(5-(9-(((9H-fluoren-9-yl)methoxy)carbonyl)-3-oxa-9-azabicyclo[3.3.1]non-6-en-7-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-5-bromo-1-ethyl-1H-indol-3-yl)-2,2-dimethylpropyl)1,2-di-tert-butyl(3S)-tetrahydropyridazine-1,2,3-tricarboxylate (6.63 g,90% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd for C₆₀H₇₂BrN₅O₁₀1101.5 & 1103.4; found 1102.4 & 1104.4.

Step 3.

To a mixture of3-(3-(2-(5-(9-(((9H-fluoren-9-yl)methoxy)carbonyl)-3-oxa-9-azabicyclo[3.3.1]non-6-en-7-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-5-bromo-1-ethyl-1H-indol-3-yl)-2,2-dimethylpropyl)1,2-di-tert-butyl(3S)-tetrahydropyridazine-1,2,3-tricarboxylate(6.4 g, 5.8 mmol) and bis(pinacolato)diboron (2.21 g, 8.7 mmol) intoluene (25 mL) under an atmosphere of N₂ was added AcOK (1.42 g, 14.5mmol) and Pd(dppf)Cl₂CH₂Cl₂ (0.47 g, 0.58 mmol) in portions. The mixturewas heated to 80° C. and stirred for 3 h, then concentrated underreduced pressure and the residue was purified by silica gel columnchromatography to give3-(3-(2-(5-(9-(((9H-fluoren-9-yl)methoxy)carbonyl)-3-oxa-9-azabicyclo[3.3.1]non-6-en-7-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-1-ethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)-2,2-dimethylpropyl)1,2-di-tert-butyl (3S)-tetrahydropyridazine-1,2,3-tricarboxylate (4.9 g,73% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd for C₆₆H₈₄BN₅O₁₂1149.6; found 1150.8.

Step 4.

To3-(3-(2-(5-(9-(((9H-fluoren-9-yl)methoxy)carbonyl)-3-oxa-9-azabicyclo[3.3.1]non-6-en-7-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-1-ethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)-2,2-dimethylpropyl)1,2-di-tert-butyl (3S)-tetrahydropyridazine-1,2,3-tricarboxylate (4.9 g,4.3 mmol) was added HCl in 1,4-dioxane (15 mL) at 0° C. The mixture waswarmed to room temperature and stirred for 5 h, then concentrated underreduced pressure to give(9H-fluoren-9-yl)methyl-7-(5-(1-ethyl-3-(3-(((S)-hexahydropyridazine-3-carbonyl)oxy)-2,2-dimethylpropyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-2-yl)-6-((S)-1-methoxyethyl)pyridin-3-yl)-3-oxa-9-azabicyclo[3.3.1]non-6-ene-9-carboxylate(4.9 g) as a solid, that was used directly in the next step withoutfurther purification. LCMS (ESI): m/z [M+H]⁺ calc'd for C₅₆H₆₈BN₅O₈949.5; found 950.5.

Step 5.

To a mixture of(9H-fluoren-9-yl)methyl-7-(5-(1-ethyl-3-(3-(((S)-hexahydropyridazine-3-carbonyl)oxy)-2,2-dimethylpropyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-2-yl)-6-((S)-1-methoxyethyl)pyridin-3-yl)-3-oxa-9-azabicyclo[3.3.1]non-6-ene-9-carboxylate(4.8 g, 5.1 mmol) and(2S,3S)-3-(4-bromo-1,3-thiazol-2-yl)-2-[(tert-butoxycarbonyl)amino]-3-ethoxypropanoicacid (3.00 g, 7.6 mmol) in DCM (30 mL) at 0° C. was added DIPEA (6.53 g,50.5 mmol) and CIP (4.22 g, 15.2 mmol) in portions. The mixture waswarmed to room temperature and stirred for 3 h, then concentrated underreduced pressure and the residue was purified by silica gel columnchromatography to give (9H-fluoren-9-yl)methyl7-(5-(3-(3-(((S)-1-((2S,3S)-3-(4-bromothiazol-2-yl)-2-((tert-butoxycarbonyl)amino)-3-ethoxypropanoyl)hexahydropyridazine-3-carbonyl)oxy)-2,2-dimethylpropyl)-1-ethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-2-yl)-6-((S)-1-methoxyethyl)pyridin-3-yl)-3-oxa-9-azabicyclo[3.3.1]non-6-ene-9-carboxylate(5.5 g, 82% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₆₉H₈₅BBrN₇O₁₂S 1325.5 & 1327.5; found 1326.5 & 1328.5.

Step 6.

To a mixture of (9H-fluoren-9-yl)methyl7-(5-(3-(3-(((S)-1-((2S,3S)-3-(4-bromothiazol-2-yl)-2-((tert-butoxycarbonyl)amino)-3-ethoxypropanoyl)hexahydropyridazine-3-carbonyl)oxy)-2,2-dimethylpropyl)-1-ethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-2-yl)-6-((S)-1-methoxyethyl)pyridin-3-yl)-3-oxa-9-azabicyclo[3.3.1]non-6-ene-9-carboxylate(5.5 g, 4.1 mmol) and K₃PO₄ (2.20 g, 10.4 mmol) in toluene (12 mL),dioxane (4 mL) and H₂O (4 mL) under an atmosphere of N₂ was addedPd(dppf)Cl₂.DCM (0.34 g, 0.41 mmol) in portions. The mixture was heatedto 80° C. and stirred for 3 h, then filtered and the filter cake waswashed with EtOAc (3×10 mL) and H₂O (10 mL). The filtrate was partionedand the aqueous layer was extracted with EtOAc (3×20 mL). The combinedorganic layers were dried over anhydrous Na₂SO₄, filtered, the filtratewas concentrated under reduced pressure and the residue was purified bysilica gel column chromatography to give(9H-fluoren-9-yl)methyl-7-(5-((6³S,3S,4S,Z)-4-((tert-butoxycarbonyl)amino)-3-ethoxy-1¹-ethyl-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-12-yl)-6-((S)-1-methoxyethyl)pyridin-3-yl)-3-oxa-9-azabicyclo[3.3.1]non-6-ene-9-carboxylate(2.3 g, 49% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₆₃H₇₃N₇O₁₀S. 1119.5; found 1120.3.

Step 7.

To a mixture of(9H-fluoren-9-yl)methyl-7-(5-((6³S,3S,4S,Z)-4-((tert-butoxycarbonyl)amino)-3-ethoxy-1¹-ethyl-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-12-yl)-6-((S)-1-methoxyethyl)pyridin-3-yl)-3-oxa-9-azabicyclo[3.3.1]non-6-ene-9-carboxylate(2.3 g, 2.1 mmol) and piperidine (0.87 g, 10.3 mmol) in DCM (10 mL) wasstirred at room temperature for 3 h, then concentrated under reducedpressure to give tert-butyl((6³S,3S,4S,Z)-1²-(5-((1R,5S)-3-oxa-9-azabicyclo[3.3.1]non-6-en-7-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-3-ethoxy-1¹-ethyl-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(2.3 g) as an oil. LCMS (ESI): m/z [M+H]⁺ calc'd for C₄₈H₆₃N₇O₈S. 897.5;found 898.9.

Step 8.

A mixture of tert-butyl((6³S,3S,4S,Z)-1²-(5-((1R,5S)-3-oxa-9-azabicyclo[3.3.1]non-6-en-7-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-3-ethoxy-1¹-ethyl-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(2.3 g, 2.6 mmol) and Pd(OH)₂, 20% on carbon (2.30 g, 3.3 mmol) in MeOH(10 mL) was hydrogenated (balloon) at room temperature for 2 h. Themixture was filtered through a pad of Celite and the filtrate wasconcentrated under reduced pressure to give tert-butyl((6³S,3S,4S,Z)-1²-(5-((1R,5S,7s)-3-oxa-9-azabicyclo[3.3.1]nonan-7-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-3-ethoxy-1¹-ethyl-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(2.3 g) as an oil, which was used directly in the next step withoutfurther purification. LCMS (ESI): m/z [M+H]⁺ calc'd for C₄₈H₆₅N₇O₈S.899.5; found 901.0.

Step 9.

To a mixture of tert-butyl((6³S,3S,4S,Z)-1²-(5-((1R,5S,7s)-3-oxa-9-azabicyclo[3.3.1]nonan-7-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-3-ethoxy-1¹-ethyl-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(480 mg, 0.53 mmol) and 3-oxetanone (154 mg, 2.1 mmol) in MeOH (4 mL) at0° C. was added AcOH (320 mg, 5.3 mmol) dropwise. The mixture was warmedto room temperature and stirred for 30 min, then the mixture wasre-cooled to 0° C. and NaBH₃CN (101 mg, 1.6 mmol) was added in portions.The mixture was heated to 60° C. and stirred for 2 h, then concentratedunder reduced pressure and the residue was purified by silica gel columnchromatography to give tert-butyl((6³S,3S,4S,Z)-3-ethoxy-1′-ethyl-1²-(2-((S)-1-methoxyethyl)-5-((1R,5S,7s)-9-(oxetan-3-yl)-3-oxa-9-azabicyclo[3.3.1]nonan-7-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(200 mg, 39% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₅₁H₆₉N₇O₉S. 955.5; found 956.7.

Step 10.

To a mixture of tert-butyl((6³S,3S,4S,Z)-3-ethoxy-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-((1R,5S,7s)-9-(oxetan-3-yl)-3-oxa-9-azabicyclo[3.3.1]nonan-7-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(200 mg, 0.21 mmol) in DCM (2 mL) at 0° C. was added TFA (0.4 mL)dropwise. The mixture was warmed to room temperature and stirred for 1h, then concentrated under reduced pressure to give(6³S,3S,4S,Z)-4-amino-3-ethoxy-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-((1R,5S,7s)-9-(oxetan-3-yl)-3-oxa-9-azabicyclo[3.3.1]nonan-7-yl)pyridin-3-yl)-10,10-dimethyl-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-5,7-dione(220 mg) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd for C₄₆H₆₁N₇O₇S.855.4; found 856.6.

Step 11.

To a mixture of(6³S,3S,4S,Z)-4-amino-3-ethoxy-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-((1R,5S,7s)-9-(oxetan-3-yl)-3-oxa-9-azabicyclo[3.3.1]nonan-7-yl)pyridin-3-yl)-10,10-dimethyl-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-5,7-dione(220 mg, 0.26 mmol) and (1r,2R,3S)-2,3-dimethylcyclopropane-1-carboxylicacid (59 mg, 0.51 mmol) in DMF (3 mL) at 0° C. under an atmosphere of N₂was added DIPEA (332 mg, 2.57 mmol) and HATU (293 mg, 0.77 mmol) inportions. The mixture was warmed to room temperature and stirred for 2h, then purified by preparative-HPLC to give(1r,2R,3S)—N-((6³S,3S,4S,Z)-3-ethoxy-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-((1R,5S,7s)-9-(oxetan-3-yl)-3-oxa-9-azabicyclo[3.3.1]nonan-7-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2,3-dimethylcyclopropane-1-carboxamide(67 mg, 27% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₅₂H₆₉N₇O₈S. 951.5; found 952.5; ¹H NMR (400 MHz, DMSO-d₆) δ 8.79 (s,1H), 8.50 (d, J=1.6 Hz, 1H), 7.92 (s, 1H), 7.79-7.72 (m, 2H), 7.65 (d,J=9.9 Hz, 1H), 7.58 (d, J=8.7 Hz, 1H), 5.86 (d, J=9.9 Hz, 1H), 5.15 (d,J=12.3 Hz, 2H), 4.91 (s, 1H), 4.74 (s, 4H), 4.37-4.20 (m, 3H), 4.18-4.05(m, 3H), 3.92 (s, 3H), 3.57 (d, J=3.5 Hz, 5H), 3.51-3.44 (m, 7H), 3.22(s, 3H), 2.90-2.74 (m, 2H), 2.55 (s, 2H), 2.39 (s, 2H), 2.06 (d, J=12.3Hz, 1H), 1.78 (d, J=29.9 Hz, 2H), 1.56-1.47 (m, 2H), 1.37 (d, J=6.1 Hz,3H), 1.16 (t, J=7.0 Hz, 5H), 1.11-0.94 (m, 7H), 0.93-0.77 (m, 6H), 0.36(s, 3H).

Example A496. Synthesis of(1S,2S)—N-((6³S,3S,4S,Z)-3-(dimethylamino)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2-methylcyclopropane-1-carboxamide

Step 1.

To a mixture of 4-bromothiazole-2-carbaldehyde (15.0 g, 78.1 mmol) and(S)-2-methylpropane-2-sulfinamide (9.47 g, 78.1 mmol) in DCM was addedCs₂CO₃ (50.90 g, 156.2 mmol) in portions. The mixture was stirred atroom temperature for 2 h, then filtered and the filter cake was washedwith DCM (3×20 mL). The filtrate was concentrated under reduced pressureand the residue was purified by silica gel column chromatography to give(S,E)-N-((4-bromothiazol-2-yl)methylene)-2-methylpropane-2-sulfinamide(26 g, 97% yield) as an oil. LCMS (ESI): m/z [M+H]⁺ calc'd forC₈H₁₁BrN₂OS₂ 294.0; found 294.8.

Step 2.

To a mixture of (R)-3,6-diethoxy-2-isopropyl-2,5-dihydropyrazine (20.57g, 96.9 mmol) in THF at −78° C. under an atmosphere of N₂ was treatedwith n-BuLi in hexanes (20.5 mL, 105.7 mmol). The mixture was stirred at−78° C. for 30 min, then(S,E)-N-((4-bromothiazol-2-yl)methylene)-2-methylpropane-2-sulfinamide(26.0 g, 88.1 mmol) was added dropwise. The mixture was stirred at −78°C. for 2 h, then warmed to 0° C. and quenched with saturated NaHCO₃. Theaqueous layer was extracted with EtOAc (3×500 mL), the combined organiclayers were concentrated under reduced pressure and the residue waspurified by silica gel column chromatography to give(S)—N—((S)-(4-bromothiazol-2-yl)((2S,5R)-3,6-diethoxy-5-isopropyl-2,5-dihydropyrazin-2-yl)methyl)-2-methylpropane-2-sulfinamide(32.0 g, 72% yield) as an oil. LCMS (ESI): m/z [M+H]⁺ calc'd forC₁₉H₃₁BrN₄O₃S₂ 506.1; found 507.0.

Step 3.

To a mixture of(S)—N—((S)-(4-bromothiazol-2-yl)((2S,5R)-3,6-diethoxy-5-isopropyl-2,5-dihydropyrazin-2-yl)methyl)-2-methylpropane-2-sulfinamide(32.0 g, 63.0 mmol) in THF (1 L) and MECN (640 mL) at 0° C. was added0.2M HCl (790 mL) dropwise. The mixture was warmed to room temperatureand stirred overnight, then quenched by the addition of saturated NaHCO₃and extracted with EtOAc (3×500 mL). The combined organic layers wereconcentrated under reduced pressure and the residue was purified bysilica gel column chromatography to give ethyl(2S,3S)-2-amino-3-(4-bromothiazol-2-yl)-3-(((S)-tert-butylsulfinyl)amino)propanoate(18.0 g, 72% yield) as an oil. LCMS (ESI): m/z [M+H]⁺ calc'd forC₁₂H₂₀BrN₃O₃S₂ 397.0; found 398.1.

Step 4.

To a mixture of ethyl(2S,3S)-2-amino-3-(4-bromothiazol-2-yl)-3-(((S)-tert-butylsulfinyl)amino)propanoate(15.0 g, 37.7 mmol) and NaHCO₃ (15.82 g, 188.3 mmol) in THF (100 mL) andH₂O (100 mL) was added FmocCl (11.69 g, 45.2 mmol) in portions. Themixture was stirred at room temperature for 2 h, then washed with H₂O(3×100 mL). The aqueous layer was extracted with EtOAc (3×100 mL), thecombined organic layers were concentrated under reduced pressure and theresidue was purified by silica gel column chromatography to give ethyl(2S,3S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-bromothiazol-2-yl)-3-(((S)-tert-butylsulfinyl)amino)propanoate(20.0 g, 86% yield) as an oil. LCMS (ESI): m/z [M+H]⁺ calc'd forC₂₇H₃₀BrN₃O₅S₂ 619.1; found 620.0.

Step 5.

A mixture of ethyl(2S,3S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-bromothiazol-2-yl)-3-(((S)-tert-butylsulfinyl)amino)propanoate(20.0 g, 32.2 mmol) and 4M HCl in MeOH (150 mL) was stirred at roomtemperature for 2 h. The mixture was concentrated under reduced pressureand the residue was purified by reverse-phase silica gel columnchromatography to give ethyl(2S,3S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-amino-3-(4-bromothiazol-2-yl)propanoate(7.5 g, 45% yield) as an oil. LCMS (ESI): m/z [M+H]⁺ calc'd forC₂₃H₂₂BrN₃O₄S. 515.1; found 516.0.

Step 6.

To a mixture of ethyl(2S,3S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-amino-3-(4-bromothiazol-2-yl)propanoate(1.2 g, 2.3 mmol) and AcOH (419 mg, 7.0 mmol) in MeOH (20 mL) was addedHCHO, 37% aqueous solution (419 mg, 13.9 mmol) and NaBH₃CN (730 mg, 11.6mmol) in portions. The mixture was stirred at room temperature for 2 h,then diluted with H₂O and extracted with EtOAc (3×200 mL). The combinedorganic layers were concentrated under reduced pressure and the residuewas purified by silica gel column chromatography to give ethyl(2S,3S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-bromothiazol-2-yl)-3-(dimethylamino)propanoate(990 mg, 78% yield) as an oil. LCMS (ESI): m/z [M+H]⁺ calc'd forC₂₅H₂₆BrN₃O₄S. 543.1; found 543.8.

Step 7.

A mixture of ethyl(2S,3S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-bromothiazol-2-yl)-3-(dimethylamino)propanoate(990 mg, 1.8 mmol) and LiOH.H₂O (174 mg, 7.3 mmol) in THF (50 mL) andH₂O (50 mL) was stirred at room temperature for 1 h, then acidified topH ˜5 with 1M HCl. The mixture was used directly in the next stepwithout further purification.

Step 8.

To the above mixture was added NaHCO₃ (764 mg, 9.1 mmol) and FmocCl (565mg, 9.1 mmol) in portions. The mixture was stirred at room temperatureovernight, then washed with H₂O (3×300 mL) and the combined aqueouslayers were extracted with EtOAc (3×30 mL). The combined organic layerswere concentrated under reduced pressure and the residue was purified bysilica gel column chromatography to give(2S,3S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-bromothiazol-2-yl)-3-(dimethylamino)propanoicacid (320 mg, 60% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₂₃H₂₂BrN₃O₄S. 515.1; found 516.0.

Step 9.

To a mixture of3-(1-ethyl-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)-2,2-dimethylpropyl(S)-hexahydropyridazine-3-carboxylate (450 mg, 0.74 mmol) and DIPEA(1.60 g, 12.4 mmol) in DMF (30 mL) was added(2S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-bromothiazol-2-yl)-3-(dimethylamino)propanoicacid (320 mg, 0.62 mmol) and HATU (471 mg, 1.24 mmol) in portions. Themixture was stirred at room temperature for 1 h, then washed with H₂O(3×30 mL) and the combined aqueous layers were extracted with EtOAc(3×30 mL). The combined organic layers were concentrated under reducedpressure and the residue was purified by silica gel columnchromatography to give3-(1-ethyl-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)-2,2-dimethylpropyl-(3S)-1-((2S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-bromothiazol-2-yl)-3-(dimethylamino)propanoyl)hexahydropyridazine-3-carboxylate(450 mg, 66% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₅₇H₆₉BBrN₇O₈S. 1101.4; found 1102.5.

Step 10.

To a mixture of3-(1-ethyl-2-(2-((S)-1-methoxyethyl)pyridin-3-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)-2,2-dimethylpropyl-(3S)-1-((2S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-bromothiazol-2-yl)-3-(dimethylamino)propanoyl)hexahydropyridazine-3-carboxylate(450 mg, 0.41 mmol) and K₃PO₄(217 mg, 1.0 mmol) in toluene (9 mL),1,4-dioxane (3 mL) and H₂O (3 mL) under an atmosphere of N₂ was addedPd(dtbpf)Cl₂ (53 mg, 0.08 mmol) in portions. The mixture was heated to60° C. and stirred for 1 h, then washed with H₂O (3×20 mL) and thecombined aqueous layers extracted with EtOAc (3×20 mL). The combinedorganic layers were concentrated under reduced pressure and the residuewas purified by silica gel column chromatography to give(9H-fluoren-9-yl)methyl((6³S,4S,Z)-3-(dimethylamino)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(70 mg, 19% yield) as an oil. LCMS (ESI): m/z [M+H]⁺ calc'd forC₅₁H₅₇N₇O₆S. 895.4; found 896.3.

Step 11.

A mixture of(9H-fluoren-9-yl)methyl((6³S,4S,Z)-3-(dimethylamino)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(70 mg, 0.08 mmol) and piperidine (0.2 mL) in MECN (2 mL) was stirred atroom temperature for 1 h. The mixture was concentrated under reducedpressure to give(6³S,4S,Z)-4-amino-3-(dimethylamino)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-5,7-dione(60 mg, 71% yield) as an oil, which was used directly in the next stepwithout further purification. LCMS (ESI): m/z [M+H]⁺ calc'd forC₃₆H₄₇N₇O₄S. 673.3; found 674.1.

Step 12.

To a mixture of(6³S,4S,Z)-4-amino-3-(dimethylamino)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-5,7-dione (60 mg, 0.09 mmol) and(1S,2S)-2-methylcyclopropane-1-carboxylic acid (9 mg, 0.09 mmol) in DMF(5 mL) was added DIPEA (230 mg, 1.78 mmol) and HATU (68 mg, 0.18 mmol)in portions. The mixture was stirred at room temperature for 1 h, thendiluted with H₂O and extracted with EtOAc (2×20 mL). The combinedorganic layers were washed with H₂O (3×20 mL), dried over anhydrousNa₂SO₄ and filtered. The filtrate was concentrated under reducedpressure and the residue was purified by preparative-HPLC to give(1S,2S)—N-((6³S,3S,4S,Z)-3-(dimethylamino)-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2-methylcyclopropane-1-carboxamide(3.6 mg, 5% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₄₁H₅₃N₇O₅S. 755.4; found 756.4; ¹H NMR (400 MHz, CD₃OD) δ 8.66-8.62 (m,1H), 8.44-8.39 (m, 1H), 7.86-7.79 (m, 1H), 7.63-7.54 (m, 2H), 7.48-7.43(m, 1H), 7.41-7.37 (m, 1H), 6.01-5.94 (m, 1H), 4.26-3.95 (m, 6H),3.94-3.81 (m, 2H), 3.47-3.38 (m, 1H), 3.07 (s, 3H), 2.80-2.72 (m, 1H),2.67-2.58 (m, 1H), 2.55-2.45 (m, 1H), 2.38-2.13 (s, 6H), 1.91 (s, 2H),1.63-1.40 (m, 3H), 1.39-1.32 (m, 3H), 1.29-1.08 (m, 7H), 1.06-0.91 (m,8H), 0.84-0.75 (m, 1H), 0.69-0.41 (m, 7H).

Example A502. Synthesis of(1r,2R,3S)—N-((6³S,3S,4S,Z)-3-ethoxy-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-((1R,5S,7r)-9-(oxetan-3-yl)-3-oxa-9-azabicyclo[3.3.1]nonan-7-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,66-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2,3-dimethylcyclopropane-1-carboxamide

Step 1.

To a mixture of tert-butyl(1R,5S)-7-oxo-3-oxa-9-azabicyclo[3.3.1]nonane-9-carboxylate (9.00 g,37.3 mmol) in MeOH (90 mL) was added 4-methoxybenzenesulfonohydrazide(9.05 g, 44.8 mmol). The mixture was heated to 50° C. and stirred for 16h, then concentrated under reduced pressure to give tert-butyl7-(2-((4-methoxyphenyl)sulfonyl)hydrazineylidene)-3-oxa-9-azabicyclo[3.3.1]nonane-9-carboxylate(15.0 g) as a solid, which was used directly in the next step withoutfurther purification. LCMS (ESI): m/z [M+H]⁺ calc'd for C₁₉H₂₇N₃O₆S.425.2; found 426.3.

Step 2.

A mixture of tert-butyl7-(2-((4-methoxyphenyl)sulfonyl)hydrazineylidene)-3-oxa-9-azabicyclo[3.3.1]nonane-9-carboxylate(12.2 g, 28.7 mmol),(S)-(5-(5-bromo-1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-1H-indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)boronicacid (21.0 g, 43.0 mmol) and Cs₂CO₃ (14.0 g, 43.0 mmol) in 1,4-dioxane(120 mL) under an atmosphere of Ar was heated to 110° C. and stirred for16 h. The mixture was concentrated under reduced pressure and theresidue was purified by silica gel column chromatography and chiral-HPLCto give tert-butyl(1R,5S,7s)-7-(5-(5-bromo-1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-1H-indol-2-yl)-6-((S)-1-methoxyethyl)pyridin-3-yl)-3-oxa-9-azabicyclo[3.3.1]nonane-9-carboxylate(3.3 g, 17% yield; RT=1.98 min) as solid and tert-butyl(1R,5S,7r)-7-(5-(5-bromo-1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-1H-indol-2-yl)-6-((S)-1-methoxyethyl)pyridin-3-yl)-3-oxa-9-azabicyclo[3.3.1]nonane-9-carboxylate(4.5 g, 23% yield, RT=2.16 min) as a solid.

Step 3.

To tert-butyl(1R,5S)-7-(5-(5-bromo-1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-1H-indol-2-yl)-6-((S)-1-methoxyethyl)pyridin-3-yl)-3-oxa-9-azabicyclo[3.3.1]nonane-9-carboxylate(4.49 g, 6.7 mmol) was added HCl in 1,4-dioxane (40 mL, 10.0 mmol) at 0°C. The mixture was warmed to room temperature and stirred for 1 h, thenconcentrated under reduced pressure to give3-(2-(5-((1R,5S,7r)-3-oxa-9-azabicyclo[3.3.1]nonan-7-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-5-bromo-1-ethyl-1H-indol-3-yl)-2,2-dimethylpropan-1-ol(4.5 g, 92% yield) as a solid, which was used directly in the next stepwithout further purification. LCMS (ESI): m/z [M+H]⁺ calc'd forC₃₀H₄₀BrN₃CO₃ 569.2; found 570.3.

Step 4.

To a mixture of3-(2-(5-((1R,5S,7r)-3-oxa-9-azabicyclo[3.3.1]nonan-7-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-5-bromo-1-ethyl-1H-indol-3-yl)-2,2-dimethylpropan-1-ol(4.49 g, 7.87 mmol) in THF (20 mL) at 0° C. was added9H-fluoren-9-ylmethyl chloroformate (3.05 g, 11.8 mmol) in aqueousNaHCO₃ (20 mL) dropwise. The mixture was warmed to room temperature andstirred for 2 h, then extracted with EtOAc (3×20 mL). The combinedorganic layers were washed with H₂O (2×20 mL), dried over anhydrousNa₂SO₄ and filtered. The filtrate was concentrated under reducedpressure and the residue was purified by silica gel columnchromatography to give (9H-fluoren-9-yl)methyl(1R,5S,7r)-7-(5-(5-bromo-1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-1H-indol-2-yl)-6-((S)-1-methoxyethyl)pyridin-3-yl)-3-oxa-9-azabicyclo[3.3.1]nonane-9-carboxylate(5.8 g, 92% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₄₅H₅₀BrN₃O₅ 793.3; found 794.3 [for ⁸¹Br].

Step 5.

To a mixture of (9H-fluoren-9-yl)methyl(1R,5S,7r)-7-(5-(5-bromo-1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-1H-indol-2-yl)-6-((S)-1-methoxyethyl)pyridin-3-yl)-3-oxa-9-azabicyclo[3.3.1]nonane-9-carboxylate(5.8 g, 7.3 mmol) and(S)-1,2-bis(tert-butoxycarbonyl)hexahydropyridazine-3-carboxylic acid(4.83 g, 14.6 mmol) in DCM (50 mL) at 0° C. was added DMAP (0.38 g, 3.1mmol) and DCC (2.55 g, 12.4 mmol) in portions. The mixture was warmed toroom temperature and stirred for 2 h, then filtered and the filter cakewas washed with DCM (3×30 mL). The filtrate was concentrated underreduced pressure and the residue was purified by silica gel columnchromatography to give 3-(3-(2-(5-((1R,5S,7r)-9-(((9H-fluoren-9-yl)methoxy)carbonyl)-3-oxa-9-azabicyclo[3.3.1]nonan-7-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-5-bromo-1-ethyl-1H-indol-3-yl)-2,2-dimethylpropyl)1,2-di-tert-butyl-(S)-tetrahydropyridazine-1,2,3-tricarboxylate(7.6 g) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd for C₆₀H₇₄BrN₅O₁₀1105.5; found 1106.4 [for ⁸¹Br].

Step 6.

To a mixture of 3-(3-(2-(5-((1R,5S,7r)-9-(((9H-fluoren-9-yl)methoxy)carbonyl)-3-oxa-9-azabicyclo[3.3.1]nonan-7-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-5-bromo-1-ethyl-1H-indol-3-yl)-2,2-dimethylpropyl)1,2-di-tert-butyl-(S)-tetrahydropyridazine-1,2,3-tricarboxylate(7.4 g, 6.7 mmol) and bis(pinacolato)diboron (8.50 g, 33.5 mmol) intoluene (70 mL) under an atmosphere of N₂ was added AcOK (2.63 g, 26.8mmol) and Pd(dppf)Cl₂CH₂Cl₂ (1.09 g, 1.34 mmol). The mixture was heatedto 80° C. and stirred for 3 h, then concentrated under reduced pressure.The residue was purified by silica gel column chromatography to give3-(3-(2-(5-((1R,5S,7r)-9-(((9H-fluoren-9-yl)methoxy)carbonyl)-3-oxa-9-azabicyclo[3.3.1]nonan-7-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-1-ethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)-2,2-dimethylpropyl)1,2-di-tert-butyl-(S)-tetrahydropyridazine-1,2,3-tricarboxylate(6.9 g, 89% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₆₆H₈₆BN₅O₁₂ 1151.6; found 1152.6.

Step 7.

To3-(3-(2-(5-((1R,5S,7r)-9-(((9H-fluoren-9-yl)methoxy)carbonyl)-3-oxa-9-azabicyclo[3.3.1]nonan-7-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-1-ethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)-2,2-dimethylpropyl)1,2-di-tert-butyl-(S)-tetrahydropyridazine-1,2,3-tricarboxylate(6.9 g, 6.0 mmol) was added HCl in 1,4-dioxane (60 mL) at 0° C. Themixture was warmed to room temperature and was stirred for 4 h, thenconcentrated under reduced pressure to give(9H-fluoren-9-yl)methyl(1R,5S,7r)-7-(5-(1-ethyl-3-(3-(((S)-hexahydropyridazine-3-carbonyl)oxy)-2,2-dimethylpropyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-2-yl)-6-((S)-1-methoxyethyl)pyridin-3-yl)-3-oxa-9-azabicyclo[3.3.1]nonane-9-carboxylate(6.47 g) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd for C₅₆H₇₀BN₅O₈951.5; found 952.6.

Step 8.

To a mixture of(9H-fluoren-9-yl)methyl(1R,5S,7r)-7-(5-(1-ethyl-3-(3-(((S)-hexahydropyridazine-3-carbonyl)oxy)-2,2-dimethylpropyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-2-yl)-6-((S)-1-methoxyethyl)pyridin-3-yl)-3-oxa-9-azabicyclo[3.3.1]nonane-9-carboxylate(6.45 g, 6.78 mmol) and(2S,3S)-3-(4-bromothiazol-2-yl)-2-((tert-butoxycarbonyl)amino)-3-ethoxypropanoicacid (3.21 g, 8.13 mmol) in DCM (60 mL) at 0° C. was added DIPEA (8.76g, 67.8 mmol) and COMU (3.48 g, 8.1 mmol) in portions. The mixture waswarmed to room temperature and stirred for 2 h, then concentrated underreduced pressure and the residue was purified by silica gel columnchromatography to give (9H-fluoren-9-yl)methyl(1R,5S,7r)-7-(5-(3-(3-(((S)-1-((2S,3S)-3-(4-bromothiazol-2-yl)-2-((tert-butoxycarbonyl)amino)-3-ethoxypropanoyl)hexahydropyridazine-3-carbonyl)oxy)-2,2-dimethylpropyl)-1-ethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-2-yl)-6-((S)-1-methoxyethyl)pyridin-3-yl)-3-oxa-9-azabicyclo[3.3.1]nonane-9-carboxylate(6.9 g, 76% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₆₉H₈₇BBrN₇O₁₂S. 1329.5; found 1330.5.

Step 9.

To a mixture of (9H-fluoren-9-yl)methyl(1R,5S,7r)-7-(5-(3-(3-(((S)-1-((2S,3S)-3-(4-bromothiazol-2-yl)-2-((tert-butoxycarbonyl)amino)-3-ethoxypropanoyl)hexahydropyridazine-3-carbonyl)oxy)-2,2-dimethylpropyl)-1-ethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-2-yl)-6-((S)-1-methoxyethyl)pyridin-3-yl)-3-oxa-9-azabicyclo[3.3.1]nonane-9-carboxylate(6.9 g, 5.2 mmol) in toluene (300 mL), 1,4-dioxane (100 mL) and H₂O (100mL) under an atmosphere of N₂ was added K₃PO₄ (3.31 g, 15.6 mmol) andPd(DtBPF)Cl₂ (0.42 g, 0.52 mmol). The mixture was heated to 80° C. andstirred for 3 h, then concentrated under reduced pressure and theresidue was purified by silica gel column chromatography to give(9H-fluoren-9-yl)methyl(1R,5S,7r)-7-(5-((63S,3S,4S,Z)-4-((tert-butoxycarbonyl)amino)-3-ethoxy-1′-ethyl-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-1²-yl)-6-((S)-1-methoxyethyl)pyridin-3-yl)-3-oxa-9-azabicyclo[3.3.1]nonane-9-carboxylate(3.8 g, 65% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₆₃H₇₅N₇O₁₀S. 1121.5; found 1123.3.

Step 10.

To a mixture of(9H-fluoren-9-yl)methyl(1R,5S,7r)-7-(5-((63S,3S,4S,Z)-4-((tert-butoxycarbonyl)amino)-3-ethoxy-1′-ethyl-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-1²-yl)-6-((S)-1-methoxyethyl)pyridin-3-yl)-3-oxa-9-azabicyclo[3.3.1]nonane-9-carboxylate(3.8 g, 3.4 mmol) in DCM (40 mL) at 0° C. was added piperidine (1.44 g,16.9 mmol) dropwise. The mixture was warmed to room temperature andstirred for 5 h, then concentrated under reduced pressure to givetert-butyl((6³S,3S,4S,Z)-1²-(5-((1R,5S,7r)-3-oxa-9-azabicyclo[3.3.1]nonan-7-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-3-ethoxy-1¹-ethyl-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(4.7 g) as a solid, which was used directly in the next step withoutfurther purification. LCMS (ESI): m/z [M+H]⁺ calc'd for C₄₈H₆₅N₇O₈S.899.5; found 900.6.

Step 11.

To a mixture of tert-butyl((6³S,3S,4S,Z)-1²-(5-((1R,5S,7r)-3-oxa-9-azabicyclo[3.3.1]nonan-7-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-3-ethoxy-1¹-ethyl-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(1.12 g, 15.6 mmol) in MeOH (2 mL) at 0° C. was added AcOH (467 mg, 7.8mmol). The mixture was stirred for 30 min, then NaBH₃CN (147 mg, 2.33mmol) was added, and the mixture was heated to 60° C. and stirred for 3h. The mixture was concentrated under reduced pressure and the residuewas purified by silica gel column chromatography to give tert-butyl((6³S,3S,4S,Z)-3-ethoxy-1′-ethyl-1²-(2-((S)-1-methoxyethyl)-5-((1R,5S,7r)-9-(oxetan-3-yl)-3-oxa-9-azabicyclo[3.3.1]nonan-7-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(251 mg, 33% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₅₁H₆₉N₇O₉S. 955.5; found 956.5.

Step 12.

To a mixture of tert-butyl((6³S,3S,4S,Z)-3-ethoxy-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-((1R,5S,7r)-9-(oxetan-3-yl)-3-oxa-9-azabicyclo[3.3.1]nonan-7-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(250 mg, 0.26 mmol) in DCM (2 mL) at 0° C. was added TFA (0.4 mL). Themixture was warmed to room temperature and stirred for 30 min, thenconcentrated under reduced pressure to give(6³S,3S,4S,Z)-4-amino-3-ethoxy-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-((1R,5S,7r)-9-(oxetan-3-yl)-3-oxa-9-azabicyclo[3.3.1]nonan-7-yl)pyridin-3-yl)-10,10-dimethyl-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-5,7-dione(310 mg) as a solid, that was used directly in the next step withoutfurther purification. LCMS (ESI): m/z [M+H]⁺ calc'd for C₄₆H₆₁N₇O₇S.855.4; found 856.5.

Step 13.

To a mixture of(6³S,3S,4S,Z)-4-amino-3-ethoxy-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-((1R,5S,7r)-9-(oxetan-3-yl)-3-oxa-9-azabicyclo[3.3.1]nonan-7-yl)pyridin-3-yl)-10,10-dimethyl-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-5,7-dione(310 mg, 0.36 mmol) and (1r,2R,3S)-2,3-dimethylcyclopropane-1-carboxylicacid (62 mg, 0.54 mmol) in DMF (3 mL) at 0° C. was added DIPEA (468 mg,3.62 mmol) and COMU (155 mg, 0.36 mmol) in portions. The mixture waswarmed to room temperature and stirred for 1 h, then purified bypreparative-HPLC to give(1r,2R,3S)—N-((6³S,3S,4S,Z)-3-ethoxy-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)-5-((1R,5S,7r)-9-(oxetan-3-yl)-3-oxa-9-azabicyclo[3.3.1]nonan-7-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,66-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2,3-dimethylcyclopropane-1-carboxamide(113 mg, 32% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₅₂H₆₉N₇O₈S. 951.5; found 952.5; ¹H NMR (300 MHz, DMSO-d₆) δ 8.66 (d,J=2.1 Hz, 1H), 8.51 (s, 1H), 7.92 (s, 1H), 7.76 (d, J=8.4 Hz, 2H), 7.61(dd, J=15.3, 9.2 Hz, 2H), 5.88 (d, J=9.9 Hz, 1H), 5.18 (d, J=11.6 Hz,1H), 4.93 (s, 1H), 4.59-4.41 (m, 3H), 4.32 (s, 2H), 4.24 (d, J=6.2 Hz,2H), 4.13 (s, 3H), 3.82 (d, J=11.0 Hz, 4H), 3.69-3.49 (m, 4H), 3.19 (s,3H), 2.84-2.74 (s, 2H), 2.64 (s, 2H), 2.10-1.94 (m, 3H), 1.76 (s, 4H),1.53 (s, 1H), 1.37 (d, J=6.1 Hz, 3H), 1.24 (s, 1H), 1.19 (d, J=7.0 Hz,3H), 1.15 (s, 2H), 1.07 (d, J=7.2 Hz, 6H), 0.93 (t, J=7.0 Hz, 3H), 0.84(s, 3H), 0.41 (s, 3H).

Example A503. Synthesis of(1r,2R,3S)—N-((6³S,3S,4S,Z)-12-(5-((1R,5S,7r)-9-cyclopropyl-3-oxa-9-azabicyclo[3.3.1]nonan-7-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-3-ethoxy-1¹-ethyl-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-11H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2,3-dimethylcyclopropane-1-carboxamide

Step 1.

To a mixture of tert-butyl((6³S,3S,4S,Z)-12-(5-((1R,5S,7r)-3-oxa-9-azabicyclo[3.3.1]nonan-7-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-3-ethoxy-1′-ethyl-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(300 mg, 0.33 mmol) and (1-ethoxycyclopropoxy)trimethylsilane (1.16 g,6.6 mmol) in MeOH (1.5 mL) at 0° C. was added AcOH (200 mg, 3.3 mmol)dropwise. The mixture was stirred at 0° C. for 30 min, then NaBH₃CN (105mg, 1.67 mmol) was added, the mixture was heated to 60° C. and stirredfor 2 h. The residue was purified by silica gel column chromatography togive tert-butyl((6³S,3S,4S,Z)-12-(5-((1R,5S,7r)-9-cyclopropyl-3-oxa-9-azabicyclo[3.3.1]nonan-7-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-3-ethoxy-1′-ethyl-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(235 mg, 75% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₅₁H₆₉N₇O₈S. 939.5; found 940.3.

Step 2.

To a mixture of tert-butyl((6³S,3S,4S,Z)-12-(5-((1R,5S,7r)-9-cyclopropyl-3-oxa-9-azabicyclo[3.3.1]nonan-7-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-3-ethoxy-1¹-ethyl-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)carbamate(230 mg, 0.25 mmol) in DCM (2 mL) at 0° C. was added TFA (0.4 mL)dropwise. The mixture was warmed to room temperature and stirred for 1h, then concentrated under reduced pressure to give(6³S,3S,4S,Z)-4-amino-1²-(5-((1R,5S,7r)-9-cyclopropyl-3-oxa-9-azabicyclo[3.3.1]nonan-7-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-3-ethoxy-1¹-ethyl-10,10-dimethyl-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-5,7-dione(340 mg) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd for C₄₆H₆₁N₇O₆S.839.4; found 840.5.

Step 3.

To a mixture of(6³S,3S,4S,Z)-4-amino-12-(5-((1R,5S,7r)-9-cyclopropyl-3-oxa-9-azabicyclo[3.3.1]nonan-7-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-3-ethoxy-1¹-ethyl-10,10-dimethyl-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-5,7-dione(200 mg, 0.24 mmol) and (1r,2R,3S)-2,3-dimethylcyclopropane-1-carboxylicacid (41 mg, 0.36 mmol) in DMF (2 mL) at 0° C. was added DIPEA (308 mg,2.4 mmol) and COMU (102 mg, 0.24 mmol) in portions. The mixture waswarmed to room temperature and stirred for 1 h, then purified bypreparative-HPLC to give(1r,2R,3S)—N-((6³S,3S,4S,Z)-12-(5-((1R,5S,7r)-9-cyclopropyl-3-oxa-9-azabicyclo[3.3.1]nonan-7-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-3-ethoxy-1¹-ethyl-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-11H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2,3-dimethylcyclopropane-1-carboxamide(78 mg, 35% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₅₂H₆₉N₇O₇S. 935.5; found 936.4; ¹H NMR (300 MHz, DMSO-d₆) δ 8.73 (m,1H), 8.50 (m, 1H), 7.93 (s, 1H), 7.82-7.53 (m, 3H), 5.88 (d, J=9.8 Hz,1H), 5.22 (d, J=9.7 Hz, 1H), 4.92 (s, 2H), 4.55-3.86 (m, 14H), 3.84-3.37(m, 7H), 3.21 (s, 3H), 2.81 (d, J=12.3 Hz, 2H), 2.32 (s, 3H), 2.02 (d,1H), 1.82 (s, 2H), 1.53 (t, J=3.9 Hz, 2H), 1.38 (d, J=6.0 Hz, 3H), 1.12(dt, J=34.8, 6.3 Hz, 13H), 0.86 (s, 7H), 0.55 (s, 3H).

Example A504. Synthesis of presumed(1S,2R,3S)—N-((6³S,3S,4S,Z)-3-ethoxy-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2-methyl-3-(pyrimidin-4-yl)cyclopropane-1-carboxamide

Step 1.

To a mixture of LiCl (0.31 g, 7.4 mmol) and pyrimidine-4-carbaldehyde(1.0 g, 9.3 mmol) in DMF at 0° C. was added DBU (1.69 g, 11.1 mmol) andtert-butyl 2-(diethoxyphosphoryl)acetate (2.80 g, 11.1 mmol) dropwise.The mixture was warmed to room temperature and stirred for 1 h at roomtemperature, then cooled to 0° C., quenched with saturated NH₄Cl andextracted with EtOAc (200 mL). The organic layer was washed with brine(3×200 mL) and concentrated under reduced pressure. The residue waspurified by silica gel column chromatography to give tert-butyl(E)-3-(pyrimidin-4-yl)acrylate (1.0 g, 52% yield) as a solid. LCMS(ESI): m/z [M+H]⁺ calc'd for C₁₁H₁₄N₂O₂ 206.1; found 207.1.

Step 2.

To a mixture ethyldiphenylsulfanium tetrafluoroborate (2.2 g, 10.2 mmol)in DME and DCM (10:1) at −60° C. under an atmosphere of N₂ was treatedwith LDA, 2M in THF (6.0 mL, 12.0 mmol) for 0.5 h. The mixture waswarmed to room temperature and tert-butyl (E)-3-(pyrimidin-4-yl)acrylate(700 mg, 3.4 mmol) was added dropwise. The mixture was stirred at roomtemperature 1 h, then quenched with saturated NH₄Cl and extracted withEtOAc (3×100 mL). The combined organic layers were concentrated underreduced pressure and the residue was purified by preparative-HPLC togive 2-methyl-3-(pyrimidin-4-yl)cyclopropane-1-carboxylic acid (160 mg,20% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd for C₁₃H₁₈N₂O₂234.1; found 235.1; ¹H NMR (300 MHz, CDCl₃) δ 9.10 (s, 1H), 8.63 (s,1H), 7.33 (s, 1H), 2.66 (dd, J=9.7, 4.7 Hz, 1H), 2.46 (dd, J=5.7, 4.7Hz, 1H), 2.06-1.78 (m, 1H), 1.47 (s, 9H), 1.13 (d, J=6.4 Hz, 3H).

A racemic mixture of the above compound was separated by chiral-HPLC togive (55 mg, single diastereomer of unknown absolute configuration,RT=6.2 min) as a solid and (61 mg, single diastereomer of unknownabsolute configuration, RT=7.3 min).

Step 3.

A mixture of tert-butyl(1S,2R,3S)-2-methyl-3-(pyrimidin-4-yl)cyclopropane-1-carboxylate (50 mg,0.21 mmol, single diastereomer of unknown absolute configuration; RT=6.2min) and TFA (5 mL, 67.3 mmol) in DCM was stirred at room temperaturefor 1 h, then concentrated under reduced pressure to give(1S,2R,3S)-2-methyl-3-(pyrimidin-4-yl)cyclopropane-1-carboxylic acid,that was used directly in the next step without further purification(single diastereomer of unknown absolute configuration). LCMS (ESI): m/z[M+H]⁺ calc'd for C₉H₁₀N₂O₂ 178.0; found 179.1.

Step 4.

To a mixture of(1S,2R,3S)-2-methyl-3-(pyrimidin-4-yl)cyclopropane-1-carboxylic acid (53mg, 0.3 mmol) and DIPEA (192 mg, 1.48 mmol) in DMF at room temperatureunder an atmosphere of N₂ was added(6³S,3S,4S,Z)-4-amino-3-ethoxy-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-5,7-dione(100 mg, 0.15 mmol) and HATU (113 mg, 0.3 mmol). The mixture was stirredat room temperature for 1 h, then diluted with EtOAc (100 mL) and washedwith brine (3×100 mL). The organic layer was concentrated under reducedpressure and the residue was purified by preparative-HPLC to give(1S,2R,3S)—N-((6³S,3S,4S,Z)-3-ethoxy-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2-methyl-3-(pyrimidin-4-yl)cyclopropane-1-carboxamide(19 mg, 15% yield; single diastereomer of unknown absoluteconfiguration) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd forC₄₅H₅₄N₈O₆S. 834.4; found 835.2; ¹H NMR (300 MHz, DMSO-d₆) δ 9.09 (s,1H), 8.82-8.71 (m, 1H), 8.66 (m, 1H), 8.51 (s, 1H), 8.23-8.22 (m, 1H),7.94 (s, 1H), 7.87-7.65 (m, 2H), 7.63-7.30 (m, 3H), 5.89-5.76 (m, 1H),5.19-5.15 (m, 1H), 4.95 (s, 1H), 4.28-4.13 (m, 5H), 3.79-3.52 (m, 7H),3.42-3.31 (m, 3H), 2.83 (s, 2H), 2.08 (s, 1H), 1.78 (s, 3H), 1.38-1.34(m, 3H), 1.31-1.20 (m, 6H), 0.88-0.78 (m, 6H), 0.41 (s, 3H).

Example A505. Synthesis of presumed(1R,2S,3R)—N-((6³S,3S,4S,Z)-3-ethoxy-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2-methyl-3-(pyrimidin-4-yl)cyclopropane-1-carboxamide

Step 1.

A racemic mixture of the starting cyclopropane was separated by chiralHPLC [condition (Column: CHIRALPAK IF, 2×25 cm, 5 μM; Mobile Phase A:Hexane (10 mM NH₃-MeOH), Mobile Phase B: IPA-HPLC; Flow rate: 20 mL/min;Gradient: 5% B to 5% B in 10 min; Wave Length: 252/220 nm; RT1 (min):6.2; RT2 (min): 7.3; Sample Solvent: MeOH:DCM=1:1; Injection Volume: 0.2mL; Number Of Runs: 11)]. Product A (single diastereomer of unknownabsolute configuration, 55 mg, RT=6.2 min) as white solid; Product B(single diastereomer of unknown absolute configuration, 61 mg, RT=7.3min).

Step 2.

A mixture of presumed tert-butyl(1R,2S,3R)-2-methyl-3-(pyrimidin-4-yl)cyclopropane-1-carboxylate (singlediastereomer of unknown absolute configuration, 50 mg, 0.21 mmol) andTFA (5 mL, 67.3 mmol) in DCM was stirred at room temperature for 1 h,then concentrated under reduced pressure to give presumed(1R,2S,3R)-2-methyl-3-(pyrimidin-4-yl)cyclopropane-1-carboxylic acid(single diastereomer of unknown absolute configuration). LCMS (ESI): m/z[M+H]⁺ calc'd for C₉H₁₀N₂O₂ 178.0; found 179.1.

Step 3.

To a mixture of(1R,2S,3R)-2-methyl-3-(pyrimidin-4-yl)cyclopropane-1-carboxylic acid (53mg, 0.3 mmol) and DIPEA (192 mg, 1.48 mmol) in DMF at room temperatureunder an atmosphere of N₂ was added(6³S,3S,4S,Z)-4-amino-3-ethoxy-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-5,7-dione(100 mg, 0.15 mmol) and HATU (113 mg, 0.3 mmol). The mixture was stirredat room temperature for 1 h, then diluted with EtOAc (100 mL) and washedwith brine (3×100 mL). The organic layer was concentrated under reducedpressure and the residue was purified by preparative-HPLC to give(1R,2S,3R)—N-((6³S,3S,4S,Z)-3-ethoxy-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2-methyl-3-(pyrimidin-4-yl)cyclopropane-1-carboxamide(single diastereomer of unknown absolute configuration, 29 mg, 19%yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc'd for C₄₅H₅₄N₈O₆S. 834.4;found 835.2; ¹H NMR (300 MHz, DMSO-d₆) δ 9.10 (s, 1H), 8.83-8.78 (m,1H), 8.71 (m, 1H), 8.54 (s, 1H), 8.29-8.24 (m, 1H), 7.98 (s, 1H),7.88-7.78 (m, 2H), 7.74-7.69 (m, 1H), 7.67-7.35 (m, 3H), 5.85-5.72 (m,1H), 5.19-5.12 (m, 1H), 4.93 (s, 1H), 4.30-4.22 (m, 5H), 3.80-3.60 (m,7H), 3.46-3.38 (m, 3H), 2.88 (s, 1H), 2.75-2.56 (m, 1H), 2.18 (s, 1H),1.88 (s, 1H), 1.71-1.56 (m, 2H), 1.48-1.24 (m, 3H), 1.28-1.22 (m, 3H),1.19-1.02 (m, 3H), 0.98-0.88 (m, 6H), 0.44 (s, 3H).

Example A506 & A507. Synthesis of presumed(1R,2S,3R)—N-((6³S,3S,4S,Z)-3-ethoxy-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2-methyl-3-(1-methyl-1H-imidazol-4-yl)cyclopropane-1-carboxamideand(1R,2S,3R)—N-((6³S,3S,4S,Z)-3-ethoxy-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2-methyl-3-(1-methyl-1H-imidazol-4-yl)cyclopropane-1-carboxamide

Step 1.

To a mixture of 1-methyl-1H-imidazole-4-carbaldehyde (1.9 g, 17.3 mmol)and LiCl (0.95 g, 22.4 mmol) in THF (10 mL) at 0° C. were addedtert-butyl 2-(diethoxyphosphoryl)acetate (5.66 g, 22.4 mmol) and DBU(2.63 g, 17.3 mmol) in portions. The mixture was allowed to warm to roomtemperature and stirred for 2 h then the mixture was washed H₂O (2×30mL). The combined aqueous layers were extracted with EtOAc (3×30 mL) andthe combined organic layers were dried and filtered. The filtrate wasconcentrated under reduced pressure and the residue was purified bysilica gel column chromatography to give tert-butyl(E)-3-(1-methyl-1H-imidazol-4-yl)acrylate (3 g, 84% yield) as an oil.LCMS (ESI): m/z [M+H]⁺ calc'd for C₁₁H₁₆N₂O₂ 208.1; found 209.1.

Step 2.

To a mixture of ethyldiphenylsulfonium tetrafluoroborate (2.18 g, 7.2mmol) in DCM:DME (1:10) at −60° C. under an atmosphere of N₂ was treatedwith 2M LDA in THF (12 mL, 24 mmol) for 30 min, followed by the additionof tert-butyl (E)-3-(1-methyl-1H-imidazol-4-yl)acrylate (500 mg, 2.4mmol) in portions. The mixture was warmed to room temperature andstirred for 2 h, then quenched with saturated NH₄Cl and extracted withEtOAc (3×50 mL). The combined organic layers were dried. The residue waspurified by preparative-HPLC to give tert-butyl(1S,2R,3S)-2-methyl-3-(1-methyl-1H-imidazol-4-yl)cyclopropane-1-carboxylate(200 mg, 35% yield of a single diastereomer of unknown absoluteconfiguration) and tert-butyl(1S,2S,3S)-2-methyl-3-(1-methyl-1H-imidazol-4-yl)cyclopropane-1-carboxylate(110 mg, 19% yield of a single diastereomer of unknown absoluteconfiguration), as an oil. LCMS (ESI): m/z [M+H]⁺ calc'd for C₁₃H₂₀N₂O₂236.2; found 236.9.

Step 3.

A mixture of presumed tert-butyl(1S,2R,3S)-2-methyl-3-(1-methyl-1H-imidazol-4-yl)cyclopropane-1-carboxylate(70 mg, 0.3 mmol) and TFA (1 mL) in DCM was stirred at room temperaturefor 2 h, then concentrated under reduced pressure to give(1S,2R,3S)-2-methyl-3-(1-methyl-1H-imidazol-4-yl)cyclopropane-1-carboxylicacid as an oil, which was used directly in the next step without furtherpurification. LCMS (ESI): m/z [M+H]⁺ calc'd for C₉H₁₂N₂O₂ 180.1; found181.3.

Step 4.

To a mixture of (6³S,3S,4S,Z)-4-amino-3-ethoxy-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-5,7-dione(122 mg, 0.18 mmol) and tert-butyl(1S,2R,3S)-2-methyl-3-(1-methyl-1H-imidazol-4-yl)cyclopropane-1-carboxylate(65 mg, 0.36 mmol) in DMF (10 mL) at room temperature was added DIPEA(467 mg, 3.6 mmol) and HATU (76 mg, 0.2 mmol) in portions. The mixturewas irradiated under microwave radiation for 2 h at room temperature.The mixture was extracted with EtOAc (2×20 mL) and the combined organiclayers were washed with H₂O (3×20 mL), dried over anhydrous Na₂SO₄ andfiltered. The filtrate was concentrated under reduced pressure and theresidue was purified by preparative-HPLC to give(1R,2S,3R)—N-((6³S,3S,4S,Z)-3-ethoxy-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2-methyl-3-(1-methyl-1H-imidazol-4-yl)cyclopropane-1-carboxamide(34 mg, 21%, diastereomer of unknown absolute configuration RT=1.00 min)as a solid and(1R,2S,3R)—N-((6³S,3S,4S,Z)-3-ethoxy-1¹-ethyl-1²-(2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-6¹,6²,6³,6⁴,6⁵,6⁶-hexahydro-1¹H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2-methyl-3-(1-methyl-1H-imidazol-4-yl)cyclopropane-1-carboxamide(36 mg, 22%, single diastereomer of unknown absolute configurationRT=1.07 min) as a solid. Data for first diastereomer (RT=1.00 min): LCMS(ESI): m/z [M+H]⁺ calc'd for C₄₅H₅₆N₈O₆S. 836.4; found 837.1; ¹H NMR(400 MHz, CD₃OD) δ 8.74-8.69 (m, 1H), 8.63-8.60 (m, 1H), 7.86-7.80 (m,1H), 7.73-7.68 (m, 1H), 7.63 (s, 1H), 7.56-7.45 (m, 3H), 6.94 (s, 1H),6.01 (s, 1H), 5.04-5.01 (m, 1H), 4.50-4.41 (m, 1H), 4.38-4.33 (m, 1H),4.29-4.20 (m, 2H), 4.15-4.06 (m, 1H), 3.76-3.73 (m, 2H), 3.72-3.68 (m,3H), 3.66-3.58 (m, 2H), 3.32-3.27 (m, 3H), 3.05-2.95 (m, 1H), 2.86-2.78(m, 1H), 2.66-2.61 (m, 1H), 2.49-2.41 (m, 1H), 2.24-2.16 (m, 1H),2.11-2.06 (m, 1H), 2.02-1.92 (m, 1H), 1.87-1.77 (m, 1H), 1.69-1.52 (m,3H), 1.46-1.41 (m, 3H), 1.38-1.15 (m, 11H), 1.04-0.81 (m, 13H), 0.50 (s,3H). Data for second diastereomer (RT=1.07 min): LCMS (ESI): m/z [M+H]⁺calc'd for C₄₅H₅₆N₈O₆S. 836.4; found 837.1; ¹H NMR (400 MHz, CD₃OD) δ8.74-8.67 (m, 1H), 8.63-8.60 (m, 1H), 7.86-7.80 (m, 1H), 7.73-7.68 (m,1H), 7.63 (s, 1H), 7.56-7.45 (m, 3H), 6.94 (s, 1H), 6.01 (s, 1H),5.04-5.01 (m, 1H), 4.50-4.41 (m, 1H), 4.38-4.33 (m, 1H), 4.29-4.20 (m,2H), 4.15-4.06 (m, 1H), 3.76-3.73 (m, 2H), 3.72-3.68 (m, 3H), 3.66-3.58(m, 2H), 3.32-3.27 (m, 3H), 3.05-2.95 (m, 1H), 2.86-2.78 (m, 1H),2.66-2.61 (m, 1H), 2.49-2.41 (m, 1H), 2.24-2.16 (m, 1H), 2.11-2.06 (m,1H), 2.02-1.92 (m, 3H), 1.87-1.77 (m, 3H), 1.68-1.53 (m, 4H), 1.46-1.41(m, 11H), 1.38-1.15 (m, 3H), 1.04-0.81 (m, 10H), 0.50 (s, 3H).

The following table of compounds were prepared using the aforementionedmethods or variations thereof, as would be known to those of skill inthe art.

TABLE 4 Exemplary Compounds Prepared by Methods of the Present InventionEx# LCMS (ESI): m/z [M + H] Found A1 799.2 A2 827.5 A3 827.5 A4 809.5 A5757.4 A6 757.4 A7 756.4 A8 784.2 A9 770.2 A10 836.2 A11 836.3 A12 738.3A13 738.3 A14 714.1 A15 853.4 A16 853.2 A17 869.2 A18 869.2 A19 880.3A20 826.2 A21 894.1 A22 840.2 A23 757.4 A24 820.2 A25 800.3 A26 786.4A27 786.4 A28 875.6 A29 865.5 A30 823.5 A31 823.5 A32 853.5 A33 853.5A34 806.8 A35 757.7 A36 811.7 A37 763.7 A38 820.4 A39 800.5 A40 882.2A41 881.9 A42 804.6 A43 804.6 A44 798.2 A45 798.2 A46 784.3 A47 796.4A48 796.4 A49 768.4 A50 818.2 A51 796.2 A52 785.2 A53 837.5 A54 837.6A55 841.5 A56 861.2 A57 833.1 A58 881.4 A59 827.2 A60 776.7 A61 776.7A62 776.7 A63 776.7 A64 757.7 A65 775.7 A66 713.6 A67 769.7 A68 749.7A69 803.7 A70 803.7 A71 770.7 A72 770.8 A73 790.8 A74 790.8 A75 787.7A76 787.8 A77 782.8 A78 777.8 A79 777.8 A80 776.7 A81 776.37 A82 771.8A83 771.8 A84 770 A85 758.7 A86 757.7 A87 755.7 A88 738.7 A89 738.7 A90741.7 A91 725.7 A92 738.7 A93 724.7 A94 743.7 A95 729.8 A96 759.5 A97759.4 A98 867.9 A99 874.1 A100 815.9 A101 768.5 A102 743.4 A103 756.5A104 771.5 A105 801.6 A106 771.2 A107 797.6 A108 724.4 A109 724.4 A110800.4 A111 784.4 A112 784.4 A113 796.4 A114 784.4 A115 791.4 A116 791.4A117 743.4 A118 825.5 A119 825.5 A120 853.5 A121 855.5 A122 811.5 A123811.6 A124 865.5 A125 724.3 A126 867.4 A127 813.5 A128 713.2 A129 742.3A130 742.3 A131 757.3 A132 772.4 A133 738.3 A134 738.3 A135 836.3 A136836.2 A137 770.2 A138 784.2 A139 756.4 A140 757.4 A141 757.4 A142 770.4A143 809.5 A144 827.5 A145 827.5 A146 400.4 A147 835.5 A148 835.5 A149862.6 A150 862.5 A151 804 A152 804.5 A153 818.5 A154 818.5 A155 756.4A156 799.4 A157 851.6 A158 771 A159 812.5 A160 850.6 A161 892.6 A162743.4 A163 856.5 A164 711.4 A165 795.8 A166 875.6 A167 823.5 A168 837.7A169 825.6 A170 822.5 A171 822.7 A172 822.5 A173 869.3 A174 917.7 A175917.7 A176 746.4 A177 728.2 A178 730.2 A179 852.4 A180 850.5 A181 886.6A182 829 A183 829 A184 852.1 A185 727.2 A186 727.2 A187 924.6 A188 924.6A189 856.6 A190 765.7 A191 756.8 A192 760.8 A193 790.8 A194 742.7 A195742.6 A196 742.4 A197 742.9 A198 783 A199 864.6 A200 864.5 A201 904.7A202 891.7 A203 898.9 A204 790.4 A205 823.7 A206 823.8 A207 841.6 A208841.6 A209 853.8 A210 764.4 A211 764.3 A212 772.3 A213 861.6 A214 792.8A215 911.7 A216 783.6 A217 772.7 A218 758.4 A219 757.9 A220 839.7 A221847.3 A222 847.3 A223 837 A224 772.6 A225 757.6 A226 800.6 A227 727.3A228 870.7 A229 810.7 A230 839.9 A231 867.7 A232 773.3 A233 727.3 A234727.4 A235 852.6 A236 792.6 A237 852.9 A238 852.8 A239 850.6 A240 906.6A241 797.6 A242 829.6 A243 898.7 A244 912.7 A245 756.2 A246 769.6 A247811.6 A248 866.6 A249 866.7 A250 866.8 A251 891.4 A252 825.7 A253 825.5A254 825.6 A255 825.8 A256 770.3 A257 883.8 A258 854.7 A259 853.5 A260890.5 A261 873.6 A262 873.6 A263 837.3 A264 881.7 A265 865.3 A266 865.5A267 865.1 A268 813.7 A269 867.4 A270 772.5 A271 913.7 A272 899.7 A273867.5 A274 851.7 A275 811.7 A276 891.3 A277 907.5 A278 897.6 A279 843.8A280 856.7 A281 857.7 A282 965.6 A283 965.5 A284 805.4 A285 768.5 A286850.9 A287 909.6 A288 925.8 A289 855.6 A290 855.6 A291 755.2 A292 755.2A293 811.8 A294 793.6 A295 384.9 A296 770.6 A297 899.7 A298 906.8 A299905.5 A300 865.5 A301 869.6 A302 869.4 A303 785.5 A304 935.4 A305 901A306 901 A307 757.4 A308 741.4 A309 771.4 A310 804.7 A311 889.7 A312910.5 A313 752.4 A314 887.4 A315 887.5 A316 869.5 A317 869.5 A318 839.7A319 839.6 A320 783.8 A321 841.8 A322 785.3 A323 799.4 A324 979.6 A325755.1 A326 755.1 A327 855.9 A328 853.6 A329 763 A330 763 A331 855.5 A332854.5 A333 885.9 A334 813.5 A335 870.7 A336 785.4 A337 769.5 A338 809.9A339 799.6 A340 739.6 A341 743.5 A342 852.7 A343 852.7 A344 820.4 A345820.4 A346 732.6 A347 732.6 A348 730.6 A349 730.3 A350 747.5 A351 745.3A352 745.3 A353 833.8 A354 870.8 A355 743.5 A356 738.5 A357 783.4 A358744.5 A359 758.6 A360 758.6 A361 728.4 A362 747.6 A363 747.5 A364 745.6A365 731.6 A366 807.4 A367 789.4 A368 746.5 A369 747.4 A370 827.4 A371842.0 A372 769.2 A373 897.4 A374 778.3 A375 806.4 A376 758.4 A377 871A378 760.5 A379 760.35 A380 760.5 A381 784.4 A382 758.5 A383 733.4 A384825.3 A385 741.2 A386 741.2 A387 807.5 A388 840.3 A389 910.7 A390 910.4A391 809.8 A392 768.6 A393 768.5 A394 770.6 A395 770.6 A396 900.9 A397886.2 A398 774.3 A399 744.6 A400 743.6 A401 918.6 A402 825.6 A403 803.5A404 803.5 A405 826.25 A406 856.7 A407 745.4 A408 881.9 A409 753.6 A410911.8 A411 885.9 A412 760.6 A413 784.3 A414 741.4 A415 769.5 A416 829.7A417 820.4 A418 866.8 A419 755.3 A420 755.3 A421 856.7 A422 739.5 A423740.4 A424 741.4 A425 763.2 A426 941.8 A427 941.3 A428 910.7 A429 899.2A430 742.3 A431 756.6 A432 756.6 A433 898.9 A434 885.4 A435 746.6 A436757.3 A437 872.3 A438 927.9 A439 820.2 A440 828.3 A441 828.2 A442 866.5A443 763.3 A444 738.1 A445 770.2 A446 756.7 A447 742.7 A448 813.6 A449910.8 A450 976.9 A451 840.7 A452 856.4 A453 838.7 A454 838.4 A455 799.4A456 813.6 A457 827.6 A458 771.2 A459 958.4 A460 928.4 A461 894.8 A462813.5 A463 820.7 A464 771.6 A465 771.6 A466 757.7 A467 856.35 A468 771.3A469 756.3 A470 745.4 A471 754.5 A472 742.2 A473 785.5 A474 799.5 A475925.5 A476 925.4 A477 870.3 A478 840.6 A479 837.6 A480 837.6 A481 932.4A482 932.4 A483 936.4 A484 952.5 A485 784.3 A486 757.5 A487 771.5 A488757.2 A489 757.25 A490 800.3 A491 790.4 A492 790.6 A493 858.3 A494 850.5A495 850.4 A496 756.4 A497 910.4 A498 790.3 A499 790.3 A500 911.8 A501911.8 A502 952.5 A503 936.4 A504 835.2 A505 835.2 A506 837.4 A507 837.3A508 854.6 A509 868.6 A510 883.7 A511 898.4 A512 897.8 A513 897.8 A514787.7 A515 883.5 A516 883.55 A517 844.3 A518 828.35 A519 770.3 A5201004.3 A521 968.9 A522 835.5 A523 821.3 A524 811.5 A525 793.4 A526 755.2A527 796.6 A528 796.3 A529 812.5 A530 812.3 A531 840.2 A532 840.2 A533824.2 A534 824.2 A535 884.3 A536 884.3 A537 884.25 A538 946.3 A5391047.6 A540 996.9 A541 997.0 A542 837.3 A543 849.3 A544 783.3 A545 756.6A546 835.7 A547 835.4 A548 884.3 A549 910.3 A550 910.7 A551 853.8 A552823.3 A553 769.3 A554 742.5 A555 953.8 A556 739.3 A557 799.2 A558 887.6A559 887.6 A560 952.5 A561 968.35 A562 968.8 A563 1011.7 A564 995.3 A5651035.7 A566 826.5 A567 826.6 A568 886.7 A569 938.8 A570 1032.4 A5711032.3 A572 828.4 A573 824.2 A574 834.3 A575 875.2 A576 824.3 A577 959.4A578 959.4 A579 868.5 A580 868.6 A581 879.25 A582 879.6 A583 866.4 A584896.2 A585 833.3 A586 887.4 A587 887.4 A588 894.4 A589 812.3 A590 812.3A591 1031.7 A592 1010.5 A593 996.6 A594 1010.4 A595 868.6 A596 858.5A597 865.2 A598 918.6 A599 868.6 A600 868.25 A601 872.3 A602 879.6 A603884.6 A604 884.4 A605 884.7 A606 880.2 A607 896.7 A608 904.3

In Vitro and In Vivo Experiments:

Potency Assay: pERK

The purpose of this assay was to measure the ability of test compoundsto inhibit K-Ras in cells. Activated K-Ras induces increasedphosphorylation of ERK at Threonine 202 and Tyrosine 204 (pERK). Thisprocedure measures a decrease in cellular pERK in response to testcompounds. The procedure described below in NCI-H358 cells is applicableto K-Ras G12C.

Note: this protocol may be executed substituting other cell lines tocharacterize inhibitors of other RAS variants, including, for example,AsPC-1 (K-Ras G12D), Capan-1 (K-Ras G12V), NCI-H1355 (K-Ras G13C), Hs766T (K-Ras Q61H), NCI-H2347 or KU-19-19 (N-Ras Q61R), or SK-MEL-30(N-Ras Q61K).

NCI-H358 cells were grown and maintained using media and proceduresrecommended by the ATCC. On the day prior to compound addition, cellswere plated in 384-well cell culture plates (40 μl/well) and grownovernight in a 37° C. 5% CO₂ incubator. Test compounds were prepared in10, 3-fold dilutions in DMSO, with a high concentration of 10 mM. On dayof assay, 40 nl of test compound was added to each well of cell cultureplate using an Echo550 liquid handler (LabCyte®). Concentrations of testcompound were tested in duplicate. After compound addition, the platesare shaken for 15 seconds at 300 rpm, centrifuged, and cells wereincubated 4 hours at 37° C. 5% CO₂. Following incubation, culture mediumwas removed and cells were washed once with phosphate buffered saline.

In some experiments, cellular pERK level was determined using theAlphaLISA SureFire Ultra p-ERK1/2 Assay Kit (PerkinElmer). Cells werelysed in 25 μl lysis buffer, with shaking at 600 RPM at roomtemperature. Lysate (10 μl) was transferred to a 384-well Opti-plate(PerkinElmer) and 5 μl acceptor mix was added. After a 2-hour incubationin the dark, 5 μl donor mix was added, plate was sealed and incubated 2hours at room temperature. Signal was read on an Envision plate reader(PerkinElmer) using standard AlphaLISA settings. Analysis of raw datawas carried out either a) in Excel (Microsoft) and Prism (GraphPad).Signal was plotted vs. the decadal logarithm of compound concentration,and IC₅₀ was determined by fitting a 4-parameter sigmoidal concentrationresponse model or b) using Genedata Screener (Genedata). Normalizedsignal was plotted vs the decadal logarithm of compounds concentration,and IC50 was determined by fitting a 4-parameter sigmoidal concentrationresponse model.

In other experiments, cellular pERK was determined by In-Cell Western.Following compound treatment, cells were washed twice with 200 μl trisbuffered saline (TBS) and fixed for 15 minutes with 150 μl 4%paraformaldehyde in TBS. Fixed cells were washed 4 times for 5 minuteswith TBS containing 0.1% Triton X-100 (TBST) and then blocked with 100μl Odyssey blocking buffer (LI-COR) for 60 minutes at room temperature.Primary antibody (pERK, CST-4370, Cell Signaling Technology) was diluted1:200 in blocking buffer, and 50 μl was added to each well and incubatedovernight at 4° C. Cells were washed 4 times for 5 minutes with TBST.Secondary antibody (IR-800CW rabbit, LI-COR, diluted 1:800) and DNAstain DRAQ5 (LI-COR, diluted 1:2000) were added and incubated 1-2 hoursat room temperature. Cells were washed 4 times for 5 minutes with TBST.Plates were scanned on a Li—COR Odyssey CLx Imager. Analysis of raw datawas carried out in Excel (Microsoft) and Prism (GraphPad). Signal wasplotted vs. the decadal logarithm of compound concentration, and IC₅₀was determined by fitting a 4-parameter sigmoidal concentration responsemodel.

All compounds A001-A608 herein exhibit an IC₅₀ of 2 μM or less in anAsPC-1 (K-Ras G12D) pERK potency assay and/or a Capan-1 (K-Ras G12V)pERK potency assay. About 40% of compounds A001-A608 were measured in aH358 K-Ras G12C context: of those that were measured, >99% had an 1C₅₀of 2 uM or under. About 40% of compounds A001-A608 were measured in aH1975 K-Ras WT context: of those that were measured, >96% had an IC50 of2 uM or under.

Disruption of B-Raf Ras-Binding Domain (BRAF^(RBD)) Interaction withK-Ras by Compounds of the Invention

Note—The following protocol describes a procedure for monitoringdisruption of K-Ras G12C (GMP-PNP) binding to BRAF^(RBD) by a compoundof the invention. This protocol may also be executed substituting otherRas proteins or nucleotides.

The purpose of this biochemical assay was to measure the ability of testcompounds to facilitate ternary complex formation between anucleotide-loaded K-Ras isoform and Cyclophilin A; the resulting ternarycomplex disrupts binding to a BRAF^(RBD) construct, inhibiting K-Rassignaling through a RAF effector. Data was reported as IC50 values.

In assay buffer containing 25 mM HEPES pH 7.3, 0.002% Tween20, 0.1% BSA,100 mM NaCl and 5 mM MgCl₂, tagless Cyclophilin A, His6-K-Ras-GMPPNP,and GST-BRAF^(RBD) were combined in a 384-well assay plate at finalconcentrations of 25 μM, 12.5 nM and 50 nM, respectively. Compound waspresent in plate wells as a 10-point 3-fold dilution series starting ata final concentration of 30 μM. After incubation at 25° C. for 3 hours,a mixture of Anti-His Eu—W1024 and anti-GST allophycocyanin was thenadded to assay sample wells at final concentrations of 10 nM and 50 nM,respectively, and the reaction incubated for an additional 1.5 hours.TR-FRET signal was read on a microplate reader (Ex 320 nm, Em 665/615nm). Compounds that facilitate disruption of a K-Ras:RAF complex wereidentified as those eliciting a decrease in the TR-FRET ratio relativeto DMSO control wells.

Over 95% of compounds A001-A608 were measured in a K-Ras G12D contextusing the Ras-Raf assay: of those that were measured, >80% had an IC50of 2 uM or under. Over 95% of compounds A001-A608 were measured in aK-Ras G12V: of those that were measured, >90% had an IC50 of 2 uM orunder. Over 95% of compounds A001-A608 were measured in a K-Ras WTcontext using the Ras-Raf assay: of those that were measured, >85% hadan IC50 of 2 uM or under. Over 95% of compounds A001-A608 were measuredin a K-Ras G12C context: of those that were measured, >90% had an IC50of 2 uM or under. Over 95% of compounds A001-A608 were measured in aK-Ras G13C context: of those that were measured, >90% had an IC50 of 2uM or under.

Determination of Cell Viability in RAS Mutant Cancer Cell LinesProtocol: CellTiter-Glo® Cell Viability Assay

Note—The following protocol describes a procedure for monitoring cellviability of KRAS mutant cancer cell lines in response to a compound ofthe invention. Other RAS isoforms may be employed, though the number ofcells to be seeded will vary based on cell line used.

The purpose of this cellular assay was to determine the effects of testcompounds on the proliferation of three human cancer cell lines(NCI-H358 (KRAS G12C), AsPC-1 (KRAS G12D), Capan-1 (KRAS G12V)) over a5-day treatment period by quantifying the amount of ATP present atendpoint using the CellTiter-Glo® 2.0 Reagent (Promega).

Cells were seeded at 250 cells/well in 40 μl of growth medium in384-well assay plates and incubated overnight in a humidified atmosphereof 5% CO₂ at 37° C. On the day of the assay, test compounds wereprepared in 9, 3-fold dilutions in DMSO, with a high concentration of 1or 10 mM as appropriate. The test compounds (40 nl) are directlydispensed to each well of cell culture plate using an Echo550 liquidhandler (LabCyte®). The plates were shaken for 15 seconds at 300 rpm,centrifuged, and incubated in a humidified atmosphere of 5% CO₂ at 37°C. for 5 days. On day 5, assay plates and their contents wereequilibrated to room temperature for approximately 30 minutes.CellTiter-Glo® 2.0 Reagent (25 μl) was added, and plate contents weremixed for 2 minutes on an orbital shaker before incubation at roomtemperature for 10 minutes. Luminescence was measured using thePerkinElmer Enspire. Data was normalized by the following: (Samplesignal/Avg. DMSO)*100. The data was fit using a four-parameter logisticfit.

Over 95% of compounds A001-A608 were measured using the CTG assay in anH358 (K-Ras G12C) cell line: of those that were measured, >90% had anIC50 of 2 uM or under. Over 80% of compounds A001-A608 were measured inan AsPC-1 (K-Ras G12D) cell line: of those that were measured, >90% hadan IC50 of 2 uM or under. Over 80% of compounds A001-A608 were measuredin a Capan-1 (K-Ras G12V) cell line: of those that were measured, >90%had an IC50 of 2 uM or under.

Compound A, a Representative Inhibitor of the Present Invention, DrivesRegressions of KRAS^(G12D) Tumors in Vivo

Methods: Effects of Compound A on tumor cell growth in vivo wereevaluated in the human pancreatic adenocarcinoma HPAC KRAS^(G12D/wt)xenograft model using female BALB/c nude mice (6-8 weeks old). Mice wereimplanted with HPAC tumor cells in PBS (3×106 cells/mouse)subcutaneously in the flank. Once tumors reached an average size of ˜150mm³, mice were randomized to treatment groups to start theadministration of test articles or vehicle. Compound A was administeredby oral gavage once every other day (po q2d). Body weight and tumorvolume (using calipers) was measured twice weekly until study endpoints.

Results: Single-agent Compound A administered at 50 mg/kg po and 100mg/kg po every other day led to complete regression of all tumors ineach group (complete regression defined as >85% tumor regression frombaseline) at the end of treatment (Day 38 after treatment started) inthe HPAC CDX model with heterozygous KRAS^(G12D) (FIG. 1A). Theanti-tumor activity of both tested doses of Compound A was statisticallysignificant compared with control group (***p<0.001, ordinary One-wayANOVA with multiple comparisons via a post-hoc Tukey's test).

Compound A, a Representative Inhibitor of the Present Invention,Regulates RAS Pathway and Drives Regressions of KRAS^(G12V) Tumors inVivo

Methods: Effects of Compound A on blood and tumor pharmacokinetics (PK),pharmacodynamics (PD), and tumor cell growth were evaluated in vivo inthe human non-small cell lung cancer (NSCLC) NCI-H441 KRAS^(G12V/wt)xenograft model using female BALB/c nude mice (6-8 weeks old). Mice wereimplanted with NCI-H441 tumor cells (2×106 cells/mouse) in 50% media,50% Matrigel, subcutaneously in the flank. For PK/PD, animals weregrouped out when tumors were ˜400 mm³ and animals were treated with asingle dose of Compound A at 10, 25 or 50 mg/kg by oral gavage. ForPK/PD n=3 measurements pertimepoint.

Once tumors reached an average size of ˜155 mm³, mice were randomized totreatment groups to start the administration of test articles orvehicle. In NCI-H441 Compound A was administered by oral gavage oncedaily (po qd) at 10 or 25 mg/kg. Body weight and tumor volume (usingcalipers) was measured twice weekly until study endpoints.

Results: Pharmacokinetics were analyzed based on total concentration(nM) of Compound A in tumors or blood, following a single oral gavagedose of Compound A at 10, 25 or 50 mg/kg, monitored through 72 hoursfollowing dose. Compound A exhibited dose-dependent exposure in bloodand tumor samples. Compound A treated at 25 mg/kg or 50 mg/kg doses wasdetectable in tumors through 72 hours following treatment (FIG. 1B). PKfrom naïve animals treated with a single dose of Compound A delivered at10 mg/kg demonstrates maximum exposure of at 2 hours (FIG. 1C). TumorDUSP6 demonstrates modulation of DUSP6, a marker of RAS pathway activityfor 72 hours following single dose administration (FIG. 1C).

Single-agent Compound A administered to NCI-H441 tumor bearing animals,treated at 10 mg/kg po qd led to regressions (reductions in tumorvolume >10% from initial) in all animals. Treated at 25 mg/kg po qd,Compound A led to complete regression of all tumors (complete regressiondefined as >85% tumor regression from baseline) at the end of treatment(Day 38 after treatment started) in the NCI-H441 CDX model withheterozygous KRAS^(G12V) (FIG. 1D, FIG. 1E). The anti-tumor activity ofCompound A was statistically significant compared with control group(***p<0.0001, ordinary One-way ANOVA with multiple comparisons via apost-hoc Tukey's test). Treatments were well tolerated by body weightmeasurements (FIG. 1F).

Compound A, a Representative Inhibitor of the Present Invention, DrivesRegressions of KRAS^(G12V) Pancreatic Ductal Adenocarcinoma andColorectal Tumors in Vivo

Methods: Effects of Compound A on tumor cell growth in vivo wereevaluated in the human pancreatic adenocarcinoma Capan-2 KRAS^(G12V/wt)and colorectal SW403 KRAS^(G12V/wt) xenograft models using female BALB/cnude mice (6-8 weeks old). Mice were implanted with Capan-2 tumor cells(4×106 (media/Matrigel) cells/mouse), or SW403 tumor cells (1×107cells/mouse) in 50% PBS, 50% Matrigel, subcutaneously in the flank. Oncetumors reached an average size of ˜160-170 mm³, mice were randomized totreatment groups to start the administration of test articles orvehicle. In Capan-2, Compound A was administered by oral gavage oncedaily (po qd) at 10 or 25 mg/kg. In SW403 CDX, Compound A wasadministered by oral gavage once daily (po qd) at 25 mg/kg or everyother day (q2d) at 50 mg/kg. Body weight and tumor volume (usingcalipers) was measured twice weekly until study endpoints.

Results: Single-agent Compound A administered to Capan-2 tumor bearinganimals, treated at 10 mg/kg po qd led to 5/8 regressions, and 25 mg·kgpo qd led to 8/8 regressions at the end of treatment (Day 38 aftertreatment started) in the Capan-2 CDX model with heterozygousKRAS^(G12V) (FIG. 2A, FIG. 2B). The anti-tumor activity of Compound Awas statistically significant compared with control group (**p<0.01,***p<0.0001, ordinary One-way ANOVA with multiple comparisons via apost-hoc Tukey's test). Treatments were well tolerated by body weightmeasurements (FIG. 2C). In SW403 CDX model, single-agent Compound Aadministered at 25 mg/kg po daily or 50 mg/kg po q2d led to significanttumor growth inhibition in all tumors through the end of treatment (Day35 after treatment started). Compound A treatment at 25 mg/kg po qddrove regressions in 8/8 tumors, and 2/8 CR's (FIG. 2D, FIG. 2E).Compound A treatment at 50 mg/kg po q2d drove regressions in 8/8 tumors.The anti-tumor activity of Compound A was statistically significantcompared with control group (***p<0.0001, ordinary One-way ANOVA withmultiple comparisons via a post-hoc Tukey's test). Treatments were welltolerated by body weight measurements (FIG. 2F).

Compound A Exhibits Potent in Vivo Inhibition of Multiple RAS-DrivenCancer Cell Lines Compounds A, B, and C Exhibit Potent in VitroInhibition of Multiple RAS-Driven Cancer Cell Lines

Methods: Potency of in vitro cell proliferation inhibition of Capan-1(KRAS^(G12V)), NCI-H358 (KRAS^(G12C)), AsPC-1 (KRAS^(G12D)), HCT116(KRAS^(G13D)), SK-MEL-30 (NRAS^(Q61K)), NCI-H1975 (EGFR^(T790M/L858R)),and/or A375 (BRAF^(V600E)) cells exposed to Compounds A, B, or C for 120hours. Cells were seeded in growth medium in 384-well assay plates andincubated overnight in a humidified atmosphere of 5% CO₂ at 37° C. Thefollowing day, cells were exposed to a 9-concentration 3-fold serialdilution of Compound A, B, or C at a starting assay concentration of 0.1μM, 1 μM, or 10 μM as appropriate. After 5 days of incubation,CellTiter-Glo® 2.0 Reagent was added to assay plates and luminescencemeasured. Data were normalized to the mean signal of DMSO-treated cells,and IC₅₀ values were estimated using a four-parameter concentrationresponse model.

Results: Compounds A, B, and C inhibited cell proliferation inRAS-driven lines (FIG. 3A, FIG. 3B, and FIG. 3C). Compound A IC50s forRAS-Driven cancer cell lines are as follows: Capan-1 (KRAS^(G12V))=1 nM,NCI-H358 (KRAS^(G12C))=1 nM, AsPC-1 (KRAS^(G12D))=3 nM, HCT116(KRAS^(G13D))=27 nM, SK-MEL-30 (NRAS^(Q61K))=13 nM, NCI-H1975(EGFR^(T790M/L858R))=1 nM. Compound B IC50s for RAS-Driven cancer celllines are as follows: Capan-1 (KRAS^(G12V))=1 nM, NCI-H358(KRAS^(G12C))=0.5 nM, AsPC-1 (KRAS^(G12D))=2 nM, HCT116 (KRAS^(G13D))=20nM, SK-MEL-30 (NRAS^(Q61K))=5 nM, NCI-H1975 (EGFR^(T790M/L858R))=1 nM.Compound C IC50s for RAS-Driven cancer cell lines are as follows:Capan-1 (KRAS^(G12V))=2 nM, NCI-H358 (KRAS^(G12C))=1 nM, AsPC-1(KRAS^(G12D))=5 nM. RAS WT-Independent cell line A375 (BRAF^(V600E)) wasnot sensitive to Compound A, B, or C treatment with IC50s >8700 nM, 4700nM, and >10000 nM, respectively.

Compound A, a Representative Inhibitor of the Present Invention, DrivesRegressions of KRAS^(G12D) Tumors in Vivo

Methods: Effects of Compound A on tumor cell growth in vivo wereevaluated in the human pancreatic adenocarcinoma HPAC KRAS^(G12D/wt) andcolorectal GP2d KRAS^(G12D/wt) xenograft models using female BALB/c nudemice (6-8 weeks old). Mice were implanted with HPAC tumor cells (3×106cells/mouse), or GP2d tumor cells (2×106 cells/mouse) in 50% PBS, 50%Matrigel, subcutaneously in the flank. Once tumors reached an averagesize of ˜150 mm³, mice were randomized to treatment groups to start theadministration of test articles or vehicle. Compound A was administeredby oral gavage once daily (po qd) at 25 mg/kg. Body weight and tumorvolume (using calipers) was measured twice weekly until study endpoints.

Results: Single-agent Compound A administered at 25 mg/kg po daily ledto complete regression of all tumors (complete regression definedas >85% tumor regression from baseline) at the end of treatment (Day 38after treatment started) in the HPAC CDX model with heterozygousKRAS^(G12D) (FIG. 4A, FIG. 4B). The anti-tumor activity of Compound Awas statistically significant compared with control group (***p<0.0001,ordinary One-way ANOVA with multiple comparisons via a post-hoc Tukey'stest). Treatments were well tolerated by body weight measurements (FIG.4C). In Gp2d CDX model, single-agent Compound A administered at 25 mg/kgpo daily led to significant tumor growth inhibition in all tumorsthrough the end of treatment (Day 35 after treatment started) (FIG. 4D,FIG. 4E). The anti-tumor activity of Compound A was statisticallysignificant compared with control group (***p<0.0001, ordinary One-wayANOVA with multiple comparisons via a post-hoc Tukey's test). Treatmentswere well tolerated by body weight measurements (FIG. 4F).

Compound A Down-Regulates Immune Checkpoint Proteins in NCI-H358, SW900,and Capan-2 Cells in Vitro

Methods: To assess the effect of Compound A on checkpoint moleculeexpression in vitro, NCI-H358, SW900 or Capan-2 cells (5e4 cells/well)were seeded in a 96-well plate and after 24 hours treated with afive-fold dilution of Compound A in the presence of 250 pg/ml IFNγ. Theplates were incubated for 48 hours at 37° C. and 5% CO₂. The cells weredetached with 0.25% Trypsin, incubated for 15 minutes in PBS containingFixable Blue Dead Cell Stain (Invitrogen) and subsequently incubatedwith FITC anti-human CD274 (PD-L1), PerCP/Cyanine5.5 anti-human CD155(PVR) and Brilliant Violet 605 anti-human CD73 (Biolegend) for 30minutes on ice. The cells were washed twice with staining buffer (PBS/2%FCS) before flow cytometric acquisition on a Cytek Auror a instrument.The analysis was performed using the SpectroFlo and FlowJo v10 software.

Results: Compound A produced a concentration-dependent 2- to 5-folddecrease of PD-L1, PVR and CD73 on NCI-H358 (FIG. 5A), SW900 (FIG. 5B),or Capan-2 (FIG. 5C) cells in vitro. Down-regulation of these proteinsis predicted to transform the immuno-suppressive tumor immunemicroenvironment in favor of anti-tumor immunity (Rothlin et al JITC2020).

Compound a, a KRAS(ON) Inhibitor Disclosed Herein, is Active Against RASOncogene Switching Mutations

FIG. 6A is a heatmap representing cellular RAS/RAF disruption assayresults regarding various KRAS mutations in the presence of differentRAS inhibitors (Compound A, a KRAS(ON) inhibitor disclosed herein, andKRAS^(G12C)(OFF) inhibitors MRTX849 (adagrasib) and AMG 510(sotorasib)).

Plasmids expressing nanoluciferase-tagged mutant KRAS4B and halo-taggedRAF1 (residues 51-149) were co-transfected into U2OS cells and incubatedfor 24 hours. Plasmids encoding the relevant mutation were generated byNew England Biolabs Q5 site-directed mutagenesis. Transfected cells werereseeded at 25000 cells/well in 96-well plates in assay media(OptiMEM+4% FBS+100 nM HaloTag NanoBRET 618 Ligand) and incubatedovernight. Promega Vivazine Nano-Glo substrate was added according tomanufacturer's instructions. Compounds were added at concentrationsranging from 0 to 10 μM and incubated for 1 hour. The luminescencesignal was measured at 460 nm and 618 nm and the BRET ratio wascalculated as the 618 nm signal divided by the 460 nm signal. The BRETratios were fit to a standard sigmoidal dose response function and theIC50 values were used to calculate the Log 2(Fold-Change) relative toKRAS^(G12C). FIG. 6B shows the IC50 value associated with each coloredbar of the heatmap.

Compound A Drives Regressions of a Syngeneic KRAS G12C Tumor Model InVivo and Synergizes with Anti-PD-1

Methods: Effects of Compound A on tumor cell growth in vivo wereevaluated in the murine syngeneic eCT26 KRAS^(G12C/G12C) ABCB1^(−/−) 120model using female Balb/c (6-8 weeks old). Mice were implanted withtumor cells (5×106 cells/mouse) in RPMI medium without supplementssubcutaneously in the upper right flank. Once tumors reached an averagesize of ˜100 mm³, mice were randomized to treatment groups to start theadministration of test articles or vehicle. Compound A was administeredby oral gavage once daily (po qd) at 25 mg/kg. InVivoMAb anti-mouse PD-1(CD279) antibody (Clone RMP1-14 from BioXCell) and InVivoMAb rat IgG2aisotype control (Clone 2A3 from BioXCell) were administered at 10 mg/kgby intraperitoneal injection biweekly (ip biw). Body weight and tumorvolume (using calipers) was measured twice weekly until study endpoints.

Results: Single-agent Compound A and the combination with anti-PD-1 ledto complete regression of all tumors at day 14 after treatment start(FIGS. 7C and 7D). The administration of Compound A was ceased at thattime in the monotherapy and combination groups. Anti-PD-1 and isotypecontrol were administered for 21 days. FIG. 7A shows the isotype controlresults. In the anti-PD-1 monotherapy group, 2/10 tumors achievedcomplete regressions (FIG. 7B). Tumor regrowth was observed in 4/10 micetreated with single agent Compound A and 0/10 mice treated with CompoundA in combination with anti-PD-1 (FIGS. 7C and 7D). Treatments were welltolerated by body weight measurements (FIG. 8). One animal in the grouptreated with Compound A monotherapy was found dead at day 14 afterdosing stop.

Compound A Modulates the Immune Tumor Microenvironment in Favor ofAnti-tumor Immunity In Vivo

Methods: eCT26 KRAS^(G12C/G12C) ABCB1^(−/−) 120 tumors were removed 24hours post the last dose after 4 days of treatment. Tumor tissue wasminced, processed with the Miltenyi Biotec Mouse Tumor Dissociation Kit,and homogenized with the GentleMACS™ Dissociator. The cell suspensionwas incubated at 4° C. for 30 minutes with Mouse BD Fc Block (Clone2.4G2 from BD Pharmingen), 10 minutes with Blue Dead Cell Stain Kit(from Invitrogen) and 30 min in cell staining buffer. Antibodies usedtargeted CD45 (Clone 30-F11 from BD Biosciences), CD19 (Clone 1D3 fromBD Biosciences), CD3ε (145-2C11 from Biolegend), CD8b (Clone H35-17.2from BD Biosciences), CD4 (Clone GK1.5 from Biolegend), F4/80 (Clone BM8from Biolegend), Ly-6G (Clone 1A8 from BD Biosciences), Ly-6C (CloneHK1.4 from Biolegend), I-A/I-E (Clone M5/114.15.2 from BD Biosciences)and CD206 (Clone C₀₆₈C₂ from Biolegend).

The eCT26 KRAS^(G12C/G12C) ABCB1^(−/−) cell line was engineered from themurine CT26 homozygous KRAS^(G12D) tumor cell line (purchased fromAmerican Type Culture Collection). Both KRAS^(G12D) alleles werereplaced with KRAS^(G12C) using CRISPR technology at Synthego. Thefollowing donor sequenceGCCTGCTGAAAATGACTGAGTATAAACTTGTGATGGTTGGAGCTTGTGGCGTAGGCAAGAGCGCCTTGACGATACAGCTAATTCAGAATCA (SEQ ID NO: 1) and guide RNA sequenceAUGGUUGGAGCUGAUGGCGU (SEQ ID NO: 2) were utilized. Additionally, theP-glycoprotein drug transporter was knocked out using the guide RNAsequences corresponding to TAAGTGGGAGCGCCACTCCA (SEQ ID NO: 3) andCCAAACACCAGCATCAAGAG (SEQ ID NO: 4), targeting the Abcb1a and Abcb1bgenes. The homozygous G12C mutation and the ABCB1 knock out wasconfirmed by Sanger sequencing in the clone I20. This clone was used forin vivo experiments.

Results: Single-agent Compound A resulted in an increase of CD8+ T-cells(FIG. 9A). Compound A also led to a decrease in M2 macrophages (FIG. 9B)and monocytic MDSCs (FIG. 9C). *p<0.05, **p<0.01 by two-tailed Student'st-test.

Compound A Exhibits Significant Anti-tumor Activity in KRAS^(G12X) TumorModels In Vivo

Methods: Effects of Compound A treatment on the growth of mutantKRAS-driven xenograft models of human NSCLC or PDAC in vivo wasevaluated in a panel of representative CDX and PDX models in femaleimmune-deficient mice. Mice were implanted with tumor cells or fragmentssubcutaneously in the flank. Once tumors reached an average size of˜150-200 mm³, mice were randomized to treatment groups to start theadministration of test articles or vehicle. Compound A was orallyadministered at 25 mg/kg once daily. Body weight and tumor volume (usingcalipers) was measured twice weekly until study endpoints. Responseswere assessed as % change from baseline tumor volume after 21-42 days oftreatment; study duration was based on controls reaching maximal tumorburden as an endpoint.

Results: Single-agent Compound A administered at 25 mg/kg po daily ledto tumor regressions in the majority of KRAS mutant models examined here(FIGS. 10A and 10B). A small minority of models did not exhibit tumorregressions but showed significant tumor growth inhibition as comparedto controls (not shown).

Compound A Significantly Extends Time to Tumor Doubling Across XenograftModels

Methods: The impact of Compound A treatment on the growth of xenografttumor models with RAS pathway aberrations (either mutations in K, H, orNRAS or upstream or downstream of RAS) in vivo was evaluated in a panelof representative CDX and PDX models in female immune-deficient mice.Mice were implanted with tumor cells or fragments subcutaneously in theflank. Once tumors reached an average size of ˜150-200 mm³, mice wererandomized to treatment groups to start the administration of testarticles or vehicle. Compound A was orally administered at 25 mg/kg oncedaily. Body weight and tumor volume (using calipers) was measured twiceweekly until study endpoints. Responses were assessed via a survivalanalysis of time to progression with progression defined % change frombaseline tumor volume within 28 days of treatment.

Results: Single-agent Compound A administered at 25 mg/kg po daily ledto a significant delay in time to tumor doubling in all the CDX and PDXxenograft models tested as compared to controls (p<0.0001 assessed via aLog-Rank test) with the population not reaching a median time in the28-day interval of this study (FIG. 11). Interestingly, when time totumor doubling was examined in the subset of models with a specificKRAS^(G12X) mutation, a further delay in time to tumor doubling modelswas observed in this subset, which was significantly differentiated fromthe overall model population (p<0.005 via a Log-Rank test). These dataindicate that tumors with KRAS^(G12X) mutation may be more susceptibleto Compound A treatment as compared to tumors with other RAS pathwayaberrations. Thus, these preclinical findings implicate KRAS^(G12X)mutation as an enrichment biomarker for patient enrollment in thecontext of RMC-Compound treatment in the clinical setting.

Compounds A, B and D Drive Regressions of KRAS^(G12V) Tumors In Vivo

Methods: The effects of compounds A, B and D on tumor cell growth invivo were evaluated in the NCI-H441 KRAS^(G12V/wt) xenograft model ofhuman non-small cell lung carcinoma using female BALB/c nude mice (6-8weeks old). Mice were implanted with tumor cells in PBS (3×10⁶cells/mouse) subcutaneously in the flank. Once tumors reached an averagesize of ˜200 mm³, mice were randomized to treatment groups to start theadministration of test articles or vehicle. Compounds were administeredorally once daily. Body weight and tumor volume (using calipers) wasmeasured twice weekly until study endpoints.

Results: Daily oral administration of each agent at 25 mg/kg led to theregression of all tumors in each treatment group at the end of treatment(Day 28 after treatment started) in this model (FIG. 12A, FIG. 12B, FIG.12C). The anti-tumor activity of each agent was statisticallysignificant compared with control group (***p<0.001, ordinary One-wayANOVA with multiple comparisons via a post-hoc Tukey's test).

Compounds A, B and D Deeply and Durably Inhibit RAS Pathway Signaling InVivo

Methods: The NCI-H441 KRAS^(G12V/wt) xenograft model of human non-smallcell lung carcinoma was used for a single-dose PKPD study. Compounds A,B and D were administered orally at 25 mg/kg. The treatment groups withsample collections at various time points were summarized in Table 5below. Tumor samples were collected to assess RAS/ERK signaling pathwaymodulation by measuring the mRNA level of human DUSP6 in qPCR assay.

TABLE 5 Summary of treatment groups, doses, and time points forsingle-dose PD study using NCI-H441 tumors Compound/group Dose/RegimenPD, n = 3/time point Vehicle control 10 ml/kg po 1 h, 24 h Compound A 25mg/kg po 1 h, 8 h, 24 h, 48 h Compound B 25 mg/kg po 1 h, 8 h, 24 h, 48h Compound D 25 mg/kg po 1 h, 8 h, 24 h, 48 h

Results: All treatments led to inhibition of DUSP6 mRNA levels in tumorsat all time points tested, indicating strong MAPK pathway modulation(FIG. 13A). And the inhibitory effects of each compound on DUSP6 mRNAlevels are durable even 48 hours after drug administration, consistentwith the durable pharmacokinetic profile observed in the blood (FIG.13B).

While the invention has been described in connection with specificembodiments thereof, it will be understood that it is capable of furthermodifications and this application is intended to cover any variations,uses, or adaptations of the invention following, in general, theprinciples of the invention and including such departures from thepresent disclosure come within known or customary practice within theart to which the invention pertains and may be applied to the essentialfeatures set forth herein.

All publications, patents and patent applications are hereinincorporated by reference in their entirety to the same extent as ifeach individual publication, patent or patent application wasspecifically and individually indicated to be incorporated by referencein its entirety.

1. A compound, or pharmaceutically acceptable salt thereof, having thestructure of Formula Ia:

wherein A is optionally substituted 3 to 6-membered cycloalkylene,optionally substituted 3 to 6-membered heterocycloalkylene, optionallysubstituted 6-membered arylene, optionally substituted 5 to 6-memberedheteroarylene, optionally substituted C₂-C₄ alkylene, or optionallysubstituted C₂-C₄ alkenylene; Y is

W is hydrogen, C₁-C₄ alkyl, optionally substituted C₁-C₃ heteroalkyl,optionally substituted 3 to 10-membered heterocycloalkyl, optionallysubstituted 3 to 10-membered cycloalkyl, optionally substituted 6 to10-membered aryl, or optionally substituted 5 to 10-membered heteroaryl;X¹ and X⁴ are each, independently, CH₂ or NH; R¹ is optionallysubstituted C₁-C₆ alkyl, optionally substituted C₁-C₆ heteroalkyl,optionally substituted 3 to 6-membered cycloalkyl, optionallysubstituted 3 to 6-membered cycloalkenyl, optionally substituted 3 to15-membered heterocycloalkyl, optionally substituted 6 to 10-memberedaryl, or optionally substituted 5 to 10-membered heteroaryl; R² ishydrogen, optionally substituted C₁-C₆ alkyl, optionally substitutedC₂-C₆ alkenyl, optionally substituted C₂-C₆ alkynyl, optionallysubstituted 3 to 6-membered cycloalkyl, optionally substituted 3 to7-membered heterocycloalkyl, optionally substituted 6-membered aryl,optionally substituted 5 or 6-membered heteroaryl; and R¹⁰ is hydrogen,hydroxy, optionally substituted C₁-C₃ alkyl, or optionally substitutedC₁-C₆ heteroalkyl.
 2. The compound of claim 1, or pharmaceuticallyacceptable salt thereof, wherein R¹ is optionally substituted 6 to10-membered aryl or optionally substituted 5 to 10-membered heteroaryl.3. The compound of claim 2, or pharmaceutically acceptable salt thereof,wherein R¹ is optionally substituted phenyl or optionally substitutedpyridine.
 4. The compound of any one of claims 1-3, or pharmaceuticallyacceptable salt thereof, wherein A is optionally substituted thiazole,optionally substituted triazole, optionally substituted morpholino,optionally substituted piperidinyl, optionally substituted pyridine, oroptionally substituted phenyl.
 5. The compound of any one of claims 1-3,or pharmaceutically acceptable salt thereof, wherein A is not anoptionally substituted phenyl or benzimidazole.
 6. The compound of claim5, or pharmaceutically acceptable salt thereof, wherein A is nothydroxyphenyl.
 7. The compound of any one of claims 1-6, orpharmaceutically acceptable salt thereof, wherein the compound is not acompound of Table
 2. 8. The compound of any one of claims 1-7, orpharmaceutically acceptable salt thereof, wherein the compound is not acompound of Table
 3. 9. The compound of any one of claims 1-8, orpharmaceutically acceptable salt thereof, wherein Y is —NHC(O)— or—NHC(O)NH—.
 10. The compound of claim 9, or pharmaceutically acceptablesalt thereof, having the structure of Formula IIa:

wherein a is 0 or
 1. 11. The compound of claim 10, or pharmaceuticallyacceptable salt thereof, having the structure of Formula II-1a:

wherein X² is N or CH; each R³ is independently selected from halogen,cyano, hydroxy, optionally substituted amine, optionally substitutedamido, optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 11-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl; and n is an integer from 1 to
 4. 12. The compound of claim11, or pharmaceutically acceptable salt thereof, having the structure ofFormula IIa-2:


13. The compound of claim 12, or pharmaceutically acceptable saltthereof, having the structure of Formula IIa-3:

wherein R⁴ and R⁵ are each independently selected from halogen, cyano,hydroxy, optionally substituted amine, optionally substituted amido,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 11-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl.
 14. The compound of claim 13, or pharmaceutically acceptablesalt thereof, having the structure of Formula IIa-4:


15. The compound of claim 14, or pharmaceutically acceptable saltthereof, having the structure of Formula IIa-5:

wherein X³ is N or CH; m is 1 or 2; R⁶, R⁷, R⁸, and R¹¹ are eachindependently selected from hydrogen, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆ heteroalkyl, optionally substituted3 to 6-membered cycloalkyl, optionally substituted 3 to 6-memberedcycloalkenyl, optionally substituted 3 to 6-membered heterocycloalkyl,optionally substituted 6 to 10-membered aryl, or optionally substituted5 to 10-membered heteroaryl; or R⁶ and R⁷ combine with the atoms towhich they are attached to form an optionally substituted 3 to8-membered cycloalkyl or an optionally substituted 3 to 8-memberedheterocycloalkyl; or R⁷ and R⁸ combine with the atoms to which they areattached to form an optionally substituted 3 to 8-memberedheterocycloalkyl; or R⁷ and R¹¹ combine with the atoms to which they areattached to form an optionally substituted 4 to 8-memberedheterocycloalkyl.
 16. The compound of claim 15, or pharmaceuticallyacceptable salt thereof, having the structure of Formula IIa-6:


17. The compound of claim 15, or pharmaceutically acceptable saltthereof, having the structure of Formula IIa-7:


18. The compound of claim 16 or 17, wherein R⁶ is methyl.
 19. Thecompound of claim 15, or pharmaceutically acceptable salt thereof,having the structure of Formula IIa-8 or Formula IIa-9:


20. The compound of claim 9, or pharmaceutically acceptable saltthereof, having the structure of Formula IIIa:

wherein a is 0 or
 1. 21. The compound of claim 20, or pharmaceuticallyacceptable salt thereof, having the structure of Formula IIIa-1:

wherein X² is N or CH; each R³ is independently selected from halogen,cyano, hydroxy, optionally substituted amine, optionally substitutedamido, optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 11-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl; and n is an integer from 1 to
 4. 22. The compound of claim21, or pharmaceutically acceptable salt thereof, having the structure ofFormula IIIa-2:


23. The compound of claim 22, or pharmaceutically acceptable saltthereof, having the structure of Formula IIIa-3:

wherein R⁴ and R⁵ are each independently selected from halogen, cyano,hydroxy, optionally substituted amine, optionally substituted amido,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 11-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl.
 24. The compound of claim 23, or pharmaceutically acceptablesalt thereof, having the structure of Formula IIIa-4:


25. The compound of claim 24, or pharmaceutically acceptable saltthereof, having the structure of Formula IIIa-5:

wherein X³ is N or CH; m is 1 or 2; R⁶, R⁷, R⁸, and R¹¹ are eachindependently selected from hydrogen, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆ heteroalkyl, optionally substituted3 to 6-membered cycloalkyl, optionally substituted 3 to 6-memberedcycloalkenyl, optionally substituted 3 to 6-membered heterocycloalkyl,optionally substituted 6 to 10-membered aryl, or optionally substituted5 to 10-membered heteroaryl; or R⁶ and R⁷ combine with the atoms towhich they are attached to form an optionally substituted 3 to8-membered cycloalkyl or an optionally substituted 3 to 8-memberedheterocycloalkyl; or R⁷ and R⁸ combine with the atoms to which they areattached to form an optionally substituted 3 to 8-memberedheterocycloalkyl; or R⁷ and R¹¹ combine with the atoms to which they areattached to form an optionally substituted 4 to 8-memberedheterocycloalkyl.
 26. The compound of claim 25, or pharmaceuticallyacceptable salt thereof, having the structure of Formula IIIa-6:


27. The compound of claim 25, or pharmaceutically acceptable saltthereof, having the structure of Formula IIIa-7:


28. The compound of claim 26 or 27, wherein R⁶ is methyl.
 29. Thecompound of claim 25, or pharmaceutically acceptable salt thereof,having the structure of Formula IIIa-8 or Formula IIIa-9:


30. The compound of claim 9, or pharmaceutically acceptable saltthereof, having the structure of Formula IVa:

wherein R⁹ is H or C₁-C₆ alkyl; and a is 0 or
 1. 31. The compound ofclaim 30, or pharmaceutically acceptable salt thereof, having thestructure of Formula IVa-1:

wherein X² is N or CH; each R³ is independently selected from halogen,cyano, hydroxy, optionally substituted amine, optionally substitutedamido, optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 11-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl; and n is an integer from 1 to
 4. 32. The compound of claim31, or pharmaceutically acceptable salt thereof, having the structure ofFormula IVa-2:


33. The compound of claim 32, or pharmaceutically acceptable saltthereof, having the structure of Formula IVa-3:

wherein R⁴ and R⁵ are each independently selected from halogen, cyano,hydroxy, optionally substituted amine, optionally substituted amido,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 11-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl.
 34. The compound of claim 33, or pharmaceutically acceptablesalt thereof, having the structure of Formula IVa-4:


35. The compound of claim 34, or pharmaceutically acceptable saltthereof, having the structure of Formula IVa-5:

wherein X³ is N or CH; m is 1 or 2; R⁶, R⁷, R⁸, and R¹¹ are eachindependently selected from hydrogen, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆ heteroalkyl, optionally substituted3 to 6-membered cycloalkyl, optionally substituted 3 to 6-memberedcycloalkenyl, optionally substituted 3 to 6-membered heterocycloalkyl,optionally substituted 6 to 10-membered aryl, or optionally substituted5 to 10-membered heteroaryl; or R⁶ and R⁷ combine with the atoms towhich they are attached to form an optionally substituted 3 to8-membered cycloalkyl or an optionally substituted 3 to 8-memberedheterocycloalkyl; or R⁷ and R⁸ combine with the atoms to which they areattached to form an optionally substituted 3 to 8-memberedheterocycloalkyl; or R⁷ and R¹¹ combine with the atoms to which they areattached to form an optionally substituted 4 to 8-memberedheterocycloalkyl.
 36. The compound of claim 35, or pharmaceuticallyacceptable salt thereof, having the structure of Formula IVa-6:


37. The compound of claim 35, or pharmaceutically acceptable saltthereof, having the structure of Formula IVa-7:


38. The compound of claim 36 or 37, wherein R⁶ is methyl.
 39. Thecompound of claim 35, or pharmaceutically acceptable salt thereof,having the structure of Formula IVa-8 or Formula IVa-9:


40. The compound of any one of claims 30-39, or pharmaceuticallyacceptable salt thereof, wherein R⁹ is methyl.
 41. The compound of anyone of claims 1-8, or pharmaceutically acceptable salt thereof, whereinY is —NHS(O)₂— or —NHS(O)₂NH—.
 42. The compound of claim 41, orpharmaceutically acceptable salt thereof, having the structure ofFormula Va:

wherein a is 0 or
 1. 43. The compound of claim 42, or pharmaceuticallyacceptable salt thereof, having the structure of Formula Va-1:

wherein X² is N or CH; each R³ is independently selected from halogen,cyano, hydroxy, optionally substituted amine, optionally substitutedamido, optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 11-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl; and n is an integer from 1 to
 4. 44. The compound of claim43, or pharmaceutically acceptable salt thereof, having the structure ofFormula Va-2:


45. The compound of claim 44, or pharmaceutically acceptable saltthereof, having the structure of Formula Va-3:

wherein R⁴ and R⁵ are each independently selected from halogen, cyano,hydroxy, optionally substituted amine, optionally substituted amido,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 11-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl.
 46. The compound of claim 45, or pharmaceutically acceptablesalt thereof, having the structure of Formula Va-4:


47. The compound of claim 46, or pharmaceutically acceptable saltthereof, having the structure of Formula Va-5:

wherein X³ is N or CH; m is 1 or 2; R⁶, R⁷, R⁸, and R¹¹ are eachindependently selected from hydrogen, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆ heteroalkyl, optionally substituted3 to 6-membered cycloalkyl, optionally substituted 3 to 6-memberedcycloalkenyl, optionally substituted 3 to 6-membered heterocycloalkyl,optionally substituted 6 to 10-membered aryl, or optionally substituted5 to 10-membered heteroaryl; or R⁶ and R⁷ combine with the atoms towhich they are attached to form an optionally substituted 3 to8-membered cycloalkyl or an optionally substituted 3 to 8-memberedheterocycloalkyl; or R⁷ and R⁸ combine with the atoms to which they areattached to form an optionally substituted 3 to 8-memberedheterocycloalkyl; or R⁷ and R¹¹ combine with the atoms to which they areattached to form an optionally substituted 4 to 8-memberedheterocycloalkyl.
 48. The compound of claim 41, or pharmaceuticallyacceptable salt thereof, having the structure of Formula VIa:

wherein a is 0 or
 1. 49. The compound of claim 48, or pharmaceuticallyacceptable salt thereof, having the structure of Formula VIa-1:

wherein X² is N or CH; each R³ is independently selected from halogen,cyano, hydroxy, optionally substituted amine, optionally substitutedamido, optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 11-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl; and n is an integer from 1 to
 4. 50. The compound of claim49, or pharmaceutically acceptable salt thereof, having the structure ofFormula VIa-2:


51. The compound of claim 50, or pharmaceutically acceptable saltthereof, having the structure of Formula VIa-3:

wherein R⁴ and R⁵ are each independently selected from halogen, cyano,hydroxy, optionally substituted amine, optionally substituted amido,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 11-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl.
 52. The compound of claim 51, or pharmaceutically acceptablesalt thereof, having the structure of Formula VIa-4:


53. The compound of claim 52, or pharmaceutically acceptable saltthereof, having the structure of Formula VIa-5:

wherein X³ is N or CH; m is 1 or 2; R⁶, R⁷, R⁸, and R¹¹ are eachindependently selected from hydrogen, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆ heteroalkyl, optionally substituted3 to 6-membered cycloalkyl, optionally substituted 3 to 6-memberedcycloalkenyl, optionally substituted 3 to 6-membered heterocycloalkyl,optionally substituted 6 to 10-membered aryl, or optionally substituted5 to 10-membered heteroaryl; or R⁶ and R⁷ combine with the atoms towhich they are attached to form an optionally substituted 3 to8-membered cycloalkyl or an optionally substituted 3 to 8-memberedheterocycloalkyl; or R⁷ and R⁸ combine with the atoms to which they areattached to form an optionally substituted 3 to 8-memberedheterocycloalkyl; or R⁷ and R¹¹ combine with the atoms to which they areattached to form an optionally substituted 4 to 8-memberedheterocycloalkyl.
 54. The compound of claim 41, or pharmaceuticallyacceptable salt thereof, having the structure of Formula VIIa:

wherein R⁹ is H or C₁-C₆ alkyl; and a is 0 or
 1. 55. The compound ofclaim 54, or pharmaceutically acceptable salt thereof, having thestructure of Formula VIIa-1:

wherein X² is N or CH; each R³ is independently selected from halogen,cyano, hydroxy, optionally substituted amine, optionally substitutedamido, optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 11-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl; and n is an integer from 1 to
 4. 56. The compound of claim55, or pharmaceutically acceptable salt thereof, having the structure ofFormula VIIa-2:


57. The compound of claim 56, or pharmaceutically acceptable saltthereof, having the structure of Formula VIIa-3:

wherein R⁴ and R⁵ are each independently selected from halogen, cyano,hydroxy, optionally substituted amine, optionally substituted amido,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 11-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl.
 58. The compound of claim 57, or pharmaceutically acceptablesalt thereof, having the structure of Formula VIIa-4:


59. The compound of claim 58, or pharmaceutically acceptable saltthereof, having the structure of Formula VIIa-5:

wherein X³ is N or CH; m is 1 or 2; R⁶, R⁷, R⁸, and R¹¹ are eachindependently selected from hydrogen, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆ heteroalkyl, optionally substituted3 to 6-membered cycloalkyl, optionally substituted 3 to 6-memberedcycloalkenyl, optionally substituted 3 to 6-membered heterocycloalkyl,optionally substituted 6 to 10-membered aryl, or optionally substituted5 to 10-membered heteroaryl; or R⁶ and R⁷ combine with the atoms towhich they are attached to form an optionally substituted 3 to8-membered cycloalkyl or an optionally substituted 3 to 8-memberedheterocycloalkyl; or R⁷ and R⁸ combine with the atoms to which they areattached to form an optionally substituted 3 to 8-memberedheterocycloalkyl; or R⁷ and R¹¹ combine with the atoms to which they areattached to form an optionally substituted 4 to 8-memberedheterocycloalkyl.
 60. The compound of any one of claims 54-59, orpharmaceutically acceptable salt thereof, wherein R⁹ is methyl.
 61. Thecompound of any one of claims 1-8, or pharmaceutically acceptable saltthereof, wherein Y is —NHS(O)— or —NHS(O)NH—.
 62. The compound of claim61, or pharmaceutically acceptable salt thereof, having the structure ofFormula VIIIa:

wherein a is 0 or
 1. 63. The compound of claim 62, or pharmaceuticallyacceptable salt thereof, having the structure of Formula VIIIa-1:

wherein X² is N or CH; each R³ is independently selected from halogen,cyano, hydroxy, optionally substituted amine, optionally substitutedamido, optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 11-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl; and n is an integer from 1 to
 4. 64. The compound of claim63, or pharmaceutically acceptable salt thereof, having the structure ofFormula VIIIa-2:


65. The compound of claim 64, or pharmaceutically acceptable saltthereof, having the structure of Formula VIIIa-3:

wherein R⁴ and R⁵ are each independently selected from halogen, cyano,hydroxy, optionally substituted amine, optionally substituted amido,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 11-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl.
 66. The compound of claim 65, or pharmaceutically acceptablesalt thereof, having the structure of Formula VIIIa-4:


67. The compound of claim 66, or pharmaceutically acceptable saltthereof, having the structure of Formula VIIIa-5:

wherein X³ is N or CH; m is 1 or 2; R⁶, R⁷, R⁸, and R¹¹ are eachindependently selected from hydrogen, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆ heteroalkyl, optionally substituted3 to 6-membered cycloalkyl, optionally substituted 3 to 6-memberedcycloalkenyl, optionally substituted 3 to 6-membered heterocycloalkyl,optionally substituted 6 to 10-membered aryl, or optionally substituted5 to 10-membered heteroaryl; or R⁶ and R⁷ combine with the atoms towhich they are attached to form an optionally substituted 3 to8-membered cycloalkyl or an optionally substituted 3 to 8-memberedheterocycloalkyl; or R⁷ and R⁸ combine with the atoms to which they areattached to form an optionally substituted 3 to 8-memberedheterocycloalkyl; or R⁷ and R¹¹ combine with the atoms to which they areattached to form an optionally substituted 4 to 8-memberedheterocycloalkyl.
 68. The compound of claim 61, or pharmaceuticallyacceptable salt thereof, having the structure of Formula IXa:

wherein a is 0 or
 1. 69. The compound of claim 68, or pharmaceuticallyacceptable salt thereof, having the structure of Formula IXa-1:

wherein X² is N or CH; each R³ is independently selected from halogen,cyano, hydroxy, optionally substituted amine, optionally substitutedamido, optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 11-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl; and n is an integer from 1 to
 4. 70. The compound of claim69, or pharmaceutically acceptable salt thereof, having the structure ofFormula IXa-2:


71. The compound of claim 70, or pharmaceutically acceptable saltthereof, having the structure of Formula IXa-3:

wherein R⁴ and R⁵ are each independently selected from halogen, cyano,hydroxy, optionally substituted amine, optionally substituted amido,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 11-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl.
 72. The compound of claim 71, or pharmaceutically acceptablesalt thereof, having the structure of Formula IXa-4:


73. The compound of claim 72, or pharmaceutically acceptable saltthereof, having the structure of Formula IXa-5:

wherein X³ is N or CH; m is 1 or 2; R⁶, R⁷, R⁸, and R¹¹ are eachindependently selected from hydrogen, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆ heteroalkyl, optionally substituted3 to 6-membered cycloalkyl, optionally substituted 3 to 6-memberedcycloalkenyl, optionally substituted 3 to 6-membered heterocycloalkyl,optionally substituted 6 to 10-membered aryl, or optionally substituted5 to 10-membered heteroaryl; or R⁶ and R⁷ combine with the atoms towhich they are attached to form an optionally substituted 3 to8-membered cycloalkyl or an optionally substituted 3 to 8-memberedheterocycloalkyl; or R⁷ and R⁸ combine with the atoms to which they areattached to form an optionally substituted 3 to 8-memberedheterocycloalkyl; or R⁷ and R¹¹ combine with the atoms to which they areattached to form an optionally substituted 4 to 8-memberedheterocycloalkyl.
 74. The compound of claim 61, or pharmaceuticallyacceptable salt thereof, having the structure of Formula Xa:

wherein R⁹ is H or C₁-C₆ alkyl; and a is 0 or
 1. 75. The compound ofclaim 74, or pharmaceutically acceptable salt thereof, having thestructure of Formula Xa-1:

wherein X² is N or CH; each R³ is independently selected from halogen,cyano, hydroxy, optionally substituted amine, optionally substitutedamido, optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 11-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl; and n is an integer from 1 to
 4. 76. The compound of claim75, or pharmaceutically acceptable salt thereof, having the structure ofFormula Xa-2:


77. The compound of claim 76, or pharmaceutically acceptable saltthereof, having the structure of Formula Xa-3:

wherein R⁴ and R⁵ are each independently selected from halogen, cyano,hydroxy, optionally substituted amine, optionally substituted amido,optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆heteroalkyl, optionally substituted 3 to 6-membered cycloalkyl,optionally substituted 3 to 6-membered cycloalkenyl, optionallysubstituted 3 to 11-membered heterocycloalkyl, optionally substituted 6to 10-membered aryl, or optionally substituted 5 to 10-memberedheteroaryl.
 78. The compound of claim 77, or pharmaceutically acceptablesalt thereof, having the structure of Formula Xa-4:


79. The compound of claim 78, or pharmaceutically acceptable saltthereof, having the structure of Formula Xa-5:

wherein X³ is N or CH; m is 1 or 2; R⁶, R⁷, R⁸, and R¹¹ are eachindependently selected from hydrogen, optionally substituted C₁-C₆alkyl, optionally substituted C₁-C₆ heteroalkyl, optionally substituted3 to 6-membered cycloalkyl, optionally substituted 3 to 6-memberedcycloalkenyl, optionally substituted 3 to 6-membered heterocycloalkyl,optionally substituted 6 to 10-membered aryl, or optionally substituted5 to 10-membered heteroaryl; or R⁶ and R⁷ combine with the atoms towhich they are attached to form an optionally substituted 3 to8-membered cycloalkyl or an optionally substituted 3 to 8-memberedheterocycloalkyl; or R⁷ and R⁸ combine with the atoms to which they areattached to form an optionally substituted 3 to 8-memberedheterocycloalkyl; or R⁷ and R¹¹ combine with the atoms to which they areattached to form an optionally substituted 4 to 8-memberedheterocycloalkyl.
 80. The compound of any one of claims 74-79, orpharmaceutically acceptable salt thereof, wherein R⁹ is methyl.
 81. Thecompound of any one of claims 10-40, 42-60, or 62-80, orpharmaceutically acceptable salt thereof, wherein a is
 0. 82. Thecompound of any one of claims 10-40, 42-60, or 62-80, orpharmaceutically acceptable salt thereof, wherein a is
 1. 83. Thecompound of any one of claims 1-82, or pharmaceutically acceptable saltthereof, wherein R² is optionally substituted C₁-C₆ alkyl.
 84. Thecompound of claim 83, or pharmaceutically acceptable salt thereof,wherein R² is selected from —CH₂CH₃ or—CH₂CFs.
 85. The compound of anyone of claims 1-84, or pharmaceutically acceptable salt thereof, whereinW is C₁-C₄ alkyl.
 86. The compound of any one of claims 1-84, orpharmaceutically acceptable salt thereof, wherein W is optionallysubstituted cyclopropyl, optionally substituted cyclobutyl, optionallysubstituted cyclopentyl, or optionally substituted cyclohexyl,optionally substituted piperidine, optionally substituted piperazine,optionally substituted pyridine, or optionally substituted phenyl. 87.The compound of any one of claims 1-84, or pharmaceutically acceptablesalt thereof, wherein W is optionally substituted 3 to 10-memberedheterocycloalkyl, optionally substituted 3 to 10-membered cycloalkyl,optionally substituted 6 to 10-membered aryl, or optionally substituted5 to 10-membered heteroaryl.
 88. The compound of any one of claims 1-84,or pharmaceutically acceptable salt thereof, wherein W is optionallysubstituted 3 to 10-membered heterocycloalkyl.
 89. The compound of claim88, or pharmaceutically acceptable salt thereof, wherein W is selectedfrom the following, or a stereoisomer thereof:


90. The compound of any one of claims 1-84, or pharmaceuticallyacceptable salt thereof, wherein W is optionally substituted 3 to10-membered cycloalkyl.
 91. The compound of claim 90, orpharmaceutically acceptable salt thereof, wherein W is selected from thefollowing, or a stereoisomer thereof:


92. The compound of any one of claims 1-84, or pharmaceuticallyacceptable salt thereof, wherein W is optionally substituted 5 to10-membered heteroaryl.
 93. The compound of claim 92, orpharmaceutically acceptable salt thereof, wherein W is selected from thefollowing, or a stereoisomer thereof:


94. The compound of any one of claims 1-84, or pharmaceuticallyacceptable salt thereof, wherein W is optionally substituted 6 to10-membered aryl.
 95. The compound of claim 94, or pharmaceuticallyacceptable salt thereof, wherein W is optionally substituted phenyl. 96.The compound of any one of claims 1-84, or pharmaceutically acceptablesalt thereof, wherein W is optionally substituted C₁-C₃ heteroalkyl. 97.The compound of claim 96, or pharmaceutically acceptable salt thereof,wherein W is selected from the following, or a stereoisomer thereof:


98. The compound of claim 85, or pharmaceutically acceptable saltthereof, wherein W is selected from the following:


99. A compound, or a pharmaceutically acceptable salt thereof, of Table1a.
 100. A pharmaceutical composition comprising a compound, or apharmaceutically acceptable salt thereof, of any one of claims 1-99 anda pharmaceutically acceptable excipient.
 101. A method of treatingcancer in a subject in need thereof, the method comprising administeringto the subject a therapeutically effective amount of a compound, or apharmaceutically acceptable salt thereof, of any one of claims 1-99 or apharmaceutical composition of claim
 100. 102. The method of claim 101,wherein the cancer comprises a Ras mutation.
 103. The method of claim102, wherein the Ras mutation is at position 12, 13 or
 61. 104. Themethod of claim 102 or 103, wherein the Ras mutation is at position 12.105. The method of claim 103, wherein the Ras mutation is at a positionselected from the group consisting of G12C, G12D, G12V, G12R, G13C,G13D, and Q61K, or a combination thereof.
 106. The method of claim 105,wherein the Ras mutation is at a position selected from the groupconsisting of G12D, G12V and G12R, or a combination thereof.
 107. Themethod of claim 106, wherein the Ras mutation is at a position selectedfrom the group consisting of G12D and G12V, or a combination thereof.108. The method of any one of claims 101-107, wherein the cancer ispancreatic cancer.
 109. The method of any one of claims 101-107, whereinthe cancer is lung cancer.
 110. The method of any one of claims 101-107,wherein the cancer is colorectal cancer.
 111. A method of treating a Rasprotein-related disorder in a subject in need thereof, the methodcomprising administering to the subject a therapeutically effectiveamount of a compound, or a pharmaceutically acceptable salt thereof, ofany one of claims 1-99 or a pharmaceutical composition of claim 100.112. A method of inhibiting a Ras protein in a cell, the methodcomprising contacting the cell with an effective amount of a compound,or a pharmaceutically acceptable salt thereof, of any one of claims 1-99or a pharmaceutical composition of claim
 100. 113. The method of claim112, wherein more than one Ras protein is inhibited in the cell. 114.The method of claim 112 or 113, wherein the cell is a cancer cell. 115.The method of claim 114, wherein the cancer cell is a pancreatic cancercell.
 116. The method of claim 114, wherein the cancer cell is a lungcancer cell.
 117. The method of claim 114, wherein the cancer cell is acolorectal cancer cell.
 118. The method of any one of claims 101-117,wherein the Ras protein is KRAS.
 119. The method or use of any one ofclaims 101-118, wherein the method further comprises administering anadditional anticancer therapy.
 120. The method of claim 119, wherein theadditional anticancer therapy is an EGFR inhibitor, a second Rasinhibitor, a SHP2 inhibitor, a SOS1 inhibitor, a Raf inhibitor, a MEKinhibitor, an ERK inhibitor, a PI3K inhibitor, a PTEN inhibitor, an AKTinhibitor, an mTORC1 inhibitor, a BRAF inhibitor, a PD-L1 inhibitor, aPD-1 inhibitor, a CDK4/6 inhibitor, a HER2 inhibitor, or a combinationthereof.
 121. The method of claim 119 or 120, wherein the additionalanticancer therapy is a SHP2 inhibitor.
 122. The method of claim 119 or120, wherein the additional anticancer therapy comprises a SHP2inhibitor and a PD-L1 inhibitor.
 123. The method of claim 119 or 120,wherein the the additional therapy comprises a second Ras inhibitor anda PD-L1 inhibitor.
 124. The method of claim 120 or 123, wherein thesecond Ras inhibitor is a KRAS^(G12C) inhibitor.
 125. The method ofclaim 123 or 124, wherein the second Ras inhibitor is a KRAS^(G12C)(ON)inhibitor.
 126. The method of claim 123 or 124, wherein the second Rasinhibitor is a KRAS^(G12C)(OFF) inhibitor.